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immunoassay (eiA) kit - BioNova

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Who We Are<br />

Arbor Assays’ focus (and our fun) is to develop and manufacture<br />

the highest quality test <strong>kit</strong>s for biomedical research.<br />

We strive for this the following ways:<br />

f Treat everyone and everything with dignity and respect<br />

f Look after our customers<br />

f Work with our vendors<br />

f Look after our employees<br />

f Contribute to our community<br />

f Give back through charitable efforts<br />

f Recycle everything we can<br />

f Look after our environment<br />

Where We CAme From<br />

In August 2007, the founders and top scientists from Assay Designs, Inc. formed Luminos. We soon realized that we would<br />

like to recognize our commitment to our location (beautiful Ann Arbor, Michigan) and to the environment around us. To show<br />

this awareness the company name was changed to Arbor Assays in the fall of 2009.<br />

our expertise<br />

The employees of Arbor Assays have more than 50 years combined experience in designing, developing, building<br />

and manufacturing both FDA in vitro diagnostic and life sciences research use only assays.<br />

One thing we NeVer compromise on is Quality. Throughout this catalog you will see the phrase “N-Cal” on<br />

several of the pages. This refers to <strong>kit</strong>s where the standard is calibrated to the US National Institute of Standards<br />

and Technology (NIST) Reference Materials. Any <strong>kit</strong> is only as good as the standard that it is referenced against.<br />

We always try to utilize the recognized National or International standard for the <strong>kit</strong>.<br />

WhAt We Do<br />

f<br />

f<br />

f<br />

Novel detection and <strong>immunoassay</strong> <strong>kit</strong> development and manufacturing—for our own products and for<br />

the Contract Assay Services we provide<br />

Contract Chemical Synthesis Routes, Hapten Labeling and Antibody Generation<br />

Specialized Sample Testing Services<br />

our CommitmeNt<br />

Our commitment is to our customers, our suppliers, our community, our environment and ourselves. Our goal is to treat<br />

everyone with respect and with courtesy as in the quote attributable to Mahatma Gandhi.<br />

“A customer is the most important visitor on our premises. He is not dependent on us. We are dependent on him. He is<br />

not an interruption in our work. He is the purpose of it. He is not an outsider in our business. He is part of it. We are not<br />

doing him a favor by serving him. He is doing us a favor by giving us an opportunity to do so.”<br />

hoW We Work With you<br />

We will give you the best technical and customer service support of any company. Our Customer and Technical Support<br />

scientists go out of their way to help you obtain the right product to achieve an accurate analytical result in a simple<br />

straightforward unambiguous experiment. That is why so many of our customers make comments like “Wow! you really are<br />

the best source of ALL information!”, JB, St. Joseph Hospital, Bangor, ME.<br />

120310<br />

Our Research The Cure Program donates<br />

a fixed amount of money for each <strong>kit</strong><br />

purchased to a single charity each year.<br />

For 2012 the selected charity is:<br />

Charcot Foundation<br />

www.ArborAssays.com | info@ArborAssays.com | Ph: +1.734.677.1774


how to order<br />

proDuCt use<br />

All of our products are for research use only; Not for Diagnostic or therapeutic use, and are intended for use by<br />

trained laboratory personnel.<br />

ViA phoNe<br />

Call us at 734-677-1774 between the hours of 8:30 am and 5:30 pm EST Monday-Friday.<br />

ViA FAx<br />

Fax orders to us 24 hours a day 7 days a week at 734-677-6860<br />

ViA emAiL<br />

Please email orders to Orders@ArborAssays.com<br />

through our Distributors<br />

Please check our web site (www.ArborAssays.com/distributors/index.asp) for our expanding list of distributors outside<br />

of the US. If you have a suggestion for a distributor in your country please send us their name and contact information.<br />

ViA mAiL<br />

Sales Order Entry<br />

Arbor Assays<br />

1514 Eisenhower Place<br />

Ann Arbor, MI 48108-3284<br />

USA<br />

terms AND CoNDitioNs<br />

We will require a valid Purchase Order from your institution or credit card. We accept Visa, MasterCard, American Express<br />

or Discover cards. For credit card orders we will need the card number, expiration date, the 3- or 4-digit security, along with<br />

the name on the card. With all orders we will also need the telephone number and email address(es) of the end user (so that<br />

shipping or important technical information can be sent) and your Accounts Payable department (if ordering via a PO).<br />

In some cases we may ask for a credit application to be filled out. Orders are typically shipped via Federal Express Standard<br />

Overnight or 2-Day AM service. If you wish the order to ship using a different carrier please let our Customer Service experts<br />

know. They will copy you on the carriers tracking information via email. You can also use your own FedEx or UPS accounts<br />

if you prefer.<br />

Our Payment Terms are strictly Net 30 days from shipment date. All payment details can be found at the bottom of the<br />

Invoice. We accept payment by check, electronic payments (ACH), and wire transfers.<br />

If we receive your order before 3:30 pm EST it will typically ship that day. In shipping thousands of orders we have never<br />

been back-ordered. However, there may be a very rare occurrence where the product will still be in manufacturing, especially<br />

with new or high volume products. If this happens we will let you know as rapidly as we can and make sure that your order<br />

is shipped as soon as possible.<br />

WArrANty iNFormAtioN<br />

Arbor Assays warrants that at the time of shipment this product is free from defects in materials and workmanship.<br />

This warranty is in lieu of any other warranty expressed or implied, including but not limited to, any implied warranty of<br />

merchantability or fitness for a particular purpose.<br />

We must be notified of any breach of this warranty within 48 hours of receipt of the product. No claim shall be honored if we<br />

are not notified within this time period, or if the product has been stored in any way other than outlined in this publication.<br />

The sole and exclusive remedy of the customer for any liability based upon this warranty is limited to the replacement of the<br />

product, or refund of the invoice price of the goods.<br />

www.ArborAssays.com | info@ArborAssays.com | Ph: +1.734.677.1774 1


2<br />

index<br />

Contact Information 2<br />

How To Place An Order 2<br />

Terms and Conditions 2<br />

Warranty Information 2<br />

Distributors 5<br />

Technical Information 46-49<br />

page Catalog #<br />

Acetylcholinesterase Fluorescent Activity Kit 6 K015-F1<br />

AChE 6<br />

ANP 7<br />

Apicidin NEW! 44/45 P011-1MG/5MG<br />

Atrial Natriuretic Peptide (ANP) EIA Kits 7 K026-H1/H5<br />

5-Azacytidine NEW! 44/45 P012-50MG/250MG<br />

BChE 8<br />

BIX-01294 NEW! 44/45 P018-5MG/25MG<br />

Blood Urea Nitrogen (BUN) Detection Kit 41 K024-H1<br />

BML-210 NEW! 44/45 P013-5MG/25MG<br />

BML-278 NEW! 44/45 P003-5MG/25MG<br />

BUN 41<br />

Butyrylcholinesterase Fluorescent Activity Kit 8 K016-F1<br />

C-646 NEW! 44/45 P014-5MG/25MG<br />

cAMP 15<br />

cAMP Inhibitors 44/45<br />

Catalase Fluorescent Activity Kit NEW! 9 K033-F1<br />

Ceruloplasmin Colorimetric Activity Kit NEW! 10 K035-H1<br />

Chemiluminescence<br />

cGMP 16<br />

cGMP Inhibitors 44/45<br />

Corticosterone CLIA Kits NEW! 11 K014-C1/C5<br />

Corticosterone EIA 11 K014-H1/H5<br />

Cortisol Affinity Resin 43 R001-500UL<br />

Cortisol EIA Kits 12 K003-H1/H1W/H5/H5W<br />

Cortisone CLIA Kits 13 K017-C1/C5<br />

Creatinine Serum Detection Kits 14 KB02-H1/H2<br />

Creatinine Low Volume 384-Well Serum Detection Kit NEW! 14 KB02-H1D<br />

Creatinine Urinary Detection Kits 14 K002-H1/H5<br />

Cyclic AMP Direct CLIA <strong>kit</strong>s 15 K019-C1/C5<br />

Cyclic AMP Direct EIA Kits 15 K019-H1/H5<br />

Cyclic GMP Direct CLIA <strong>kit</strong>s 16 K020-C1/C5<br />

Cyclic GMP Direct EIA Kits 16 K020-H1/H5<br />

Cystatin C EIA Kit 17 K012-H1<br />

L-Cysteine Monoclonal Antibody 44/45 A002-50UG<br />

www.ArborAssays.com | info@ArborAssays.com | Ph: +1.734.677.1774


page Catalog #<br />

index Continued<br />

Decitibine NEW! 44/45 P015-10MG/50MG<br />

13,14-Dihydro-15-keto-Prostaglandin F 2a<br />

DiMethyl Lysine 4 Histone H3 Polyclonal Antibody 43 A006-100UL<br />

DyLight ® 488/550 43<br />

E1Gluc 20<br />

EIA/ELISA 46<br />

Estradiol EIA Kits 18 K030-H1/H5<br />

Estrone EIA Kits NEW! 19 K031-H1/H5<br />

Estrone-3-Glucuronide (E1G) EIA Kits NEW! 20 K036-H1/H5<br />

EX-527 NEW! 44/45 P005-5MG/25MG<br />

FK-866 NEW! 44/45 P006-5MG/25Mg<br />

Formaldehyde Fluorescent Detection Kit 21 K001-F1<br />

Garcinol NEW! 44/45 P017-5MG/25MG<br />

Glutathione Colorimetric Detection Kit 22 K006-H1<br />

Glutathione Colorimetric Cuvette Detection Kits NEW! 22 K006-H1L/H1H<br />

Glutathione Fluorescent Detection Kits 22 K006-F1/F5<br />

Glutathione Monoclonal Antibody 43 A001-50UG<br />

Glutathione Monoclonal Antibody DyLight ® 488/550 43 A001F-50UG/A001T-50UG<br />

Glutathione Reductase Fluorescent Activity Kit 23 K009-F1<br />

Glutathione S-Transferase Fluorescent Activity Kit 24 K008-F1<br />

GSH 22<br />

GSSG 22<br />

GST 24<br />

H 2 O 2<br />

HDM 26<br />

Hemoglobin Colorimetric Detection Kit 25 K013-H1<br />

Hexyl-4-Pentynoic Acid (HPA) NEW! 43 P008-10MG/590MG<br />

Hgb 25<br />

Histone Demethylase Fluorescent Activity Kit 26 K010-F1<br />

Histone H3 Polyclonal Antibody 43 A004-100UL<br />

Hydrogen Peroxide Fluorescent Detection Kit NEW! 27 K034-F1<br />

IBMX NEW! 43 P019-100MG/1GM<br />

L-Cysteine Monoclonal Antibody 43 A002-50UG<br />

LSD1 Polyclonal Antibody 43 A003-100UL<br />

MonoMethyl Lysine 4 - Histone H3 Polyclonal antibody 43 A005-100UL<br />

Mouse IgG, Fc, Goat Antibody NEW! 43 A008-10MG/25MG<br />

Nitric Oxide Colorimetric Detection Kit 28 K023-H1<br />

www.ArborAssays.com | info@ArborAssays.com | Ph: +1.734.677.1774 3<br />

34<br />

27


4<br />

index Continued<br />

page Catalog #<br />

Obelin Bioluminescent Protein 42 L001-50UG/100UG<br />

OPN 29<br />

Osteopontin EIA Kit NEW! 29 K021-H1<br />

P450 Demethylating Fluorescent Activity Kit 30 K011-F1<br />

Palladium API Fluorescent Detection Kit 31 K007-F1<br />

PDG 33<br />

PGE 2<br />

PGFM EIA Kits 32 K022-H1/H5<br />

Phenylbutyrate, Sodium NEW! 44/45 P007-1GM<br />

Piceatannol NEW! 44/45 P009-5MG/25MG<br />

PKA Colorimetric Activity Kit 35 K027-H1<br />

Pregnanediol-3 Glucuronide NEW! 33 K037-H1/H5<br />

Progesterone EIA Kits 34 K025-H1/H5<br />

Prostaglandin E 2 CLIA Kits 36 K018-C1/C5<br />

Prostaglandin E 2 EIA Kits 36 K018-H1/H5<br />

Prostaglandin E 2 High Sensitivity EIA Kits 36 K018-HX1/HX5<br />

PKA 35<br />

Rabbit IgG, Fc, Goat Antibody NEW! 43 A009-10MG/25MG<br />

RBP 37<br />

Resveratrol NEW! 44/45 P002-100MG/500MG<br />

Retinol Binding Protein Serum EIA Kit 37 K004-H1<br />

Retinol Binding Protein Urinary EIA Kit 37 KU04-H1<br />

SAHA NEW! 44/45 P004-50MG/250MG<br />

Sheep IgG, Donkey Antibody NEW! 43 A010-10MG/25MG<br />

Sirtinol NEW! 44/45 P016-5MG/25MG<br />

SOD 38<br />

Sodium Valproate 44/45<br />

Superoxide Dismutase Colorimetric Activity Kit 38 K028-H1<br />

Testosterone EIA Kits NEW! 39 K032-H1/H5<br />

Thiol Fluorescent Detection Kit 40 K005-F1<br />

ThioStar ® Thiol Detection Reagent 42 L002-50UG/100UG/250UG/500UG<br />

Tranylcypromine 44/45 X042-1EA<br />

Trichostatin A NEW! 44/45 P010-1MG<br />

TriMethyl Lysine4 Histone H3 Polyclonal Antibody 43 A007-100UL<br />

Urea Nitrogen (BUN) Colorimetric Detection Kit 41 K024-H1<br />

Urinary Creatinine Detection Kits 14<br />

Urinary Retinol Binding Protein EIA Kit 37<br />

Valproate, Sodium NEW! 44/45 P001-5GM<br />

Vorinostat 44/45<br />

36<br />

www.ArborAssays.com | info@ArborAssays.com | Ph: +1.734.677.1774


international Distributors<br />

Co u n t r y Di s t r i b u t o r We b s i t e Co u n t r y Di s t r i b u t o r We b s i t e<br />

Argentina AbBcn www.Antibody.com.ar kuwait DPC Lebanon www.dpcleb.com<br />

Australia BioScientific www.biosci.com.au Lebanon DPC Lebanon www.dpcleb.com<br />

VWR au.vwr.com Luxembourg Bio-Connect www.bio-connect.nl<br />

Austria BIOTREND www.biotrend.com Gentaur www.gentaur.com<br />

bahrain DPC Lebanon www.dpcleb.com SanBio www.sanbio.nl<br />

belarus BioTest www.biotst.net malaysia Axon www.axonscientific.com<br />

belgium Bio-Connect www.bio-connect.nl mexico Bioselec www.bioselec.com.mx<br />

Gentaur www.gentaur.com Netherlands Bio-Connect www.bio-connect.nl<br />

SanBio www.sanbio.nl Gentaur www.gentaur.com<br />

brazil Life Sciences www.life-sciences.com.br SanBio www.sanbio.nl<br />

Sellex www.sellex.com New Zealand BioScientific www.biosci.com.au<br />

Canada Arbor Assays www.arborassays.com Global Science www.globalscience.co.nz<br />

Cedarlane www.cedarlanelabs.com Nigeria Egy-Chem www.egy-chem.com<br />

Chile AbCL www.antibody.cl Norway Nordic Biosite www.nordicbiosite.com<br />

China Beijing Sheng Kebo www.fanbiotech.com oman DPC Lebanon www.dpcleb.com<br />

biocoen www.biocoen.com pakistan 3A Medical 3a_medicalsys@live.com<br />

Dakewe www.dakewe.net poland LuBioSciences www.lubio.ch<br />

Shanghai Universal www.univ-bio.com portugal bioNova www.bionova.es<br />

Zhonghao (Biopcr) www.biopcr.com Labnet www.labnet.es<br />

Czechoslovakia LuBioSciences www.lubio.ch Qatar Astra Medcare www.astramedcare.com<br />

Denmark Nordic Biosite www.nordicbiosite.com DPC Lebanon www.dpcleb.com<br />

egypt DPC Lebanon www.dpcleb.com russia BioTest www.biotst.net<br />

Egy-Chem www.egy-chem.com saudi Arabia DPC Lebanon www.dpcleb.com<br />

Finland Nordic Biosite www.nordicbiosite.com Salima www.salimacorp.com<br />

France Euromedex www.euromedex.com singapore vCell Science www.vcellscience.com<br />

germany BIOTREND www.biotrend.com south Africa Biocom Biotech www.biocombiotech.co.za<br />

hungary LuBioSciences www.lubio.ch spain bioNova www.bionova.es<br />

india Biogenuix www.biogenuix.com sudan Egy-Chem www.egy-chem.com<br />

indonesia PT Genetika www.ptgenetika.com sweden Nordic Biosite www.nordicbiosite.com<br />

iraq DPC Lebanon www.dpcleb.com switzerland BIOTREND www.biotrend.com<br />

ireland Bioquote www.bioquote.com LuBioSciences www.lubio.ch<br />

Tebu-Bio www.tebu-bio.com taiwan Level www.level.com.tw<br />

israel Enco www.enco.co.il thailand Bio-Active www.bio-active.co.th<br />

ms-biotec www.ms-biotec.co.il turkey Oksante www.oksante.com.tr<br />

italy TEMA RICERCA www.temaricerca.com uk Bioquote www.bioquote.com<br />

Japan Funakoshi www.funakoshi.co.jp Tebu-Bio www.tebu-bio.com<br />

Nacalai www.nacalai.co.jp uAe DPC Lebanon www.dpcleb.com<br />

Jordan DPC Lebanon www.dpcleb.com ukraine BioTest www.biotst.net<br />

kazakhstan BioTest www.biotst.net us Arbor Assays www.ArborAssays.com<br />

korea Dong In Biotech www.donginbio.com Cedarlane www.cedarlanelabs.com<br />

www.ArborAssays.com | info@ArborAssays.com | Ph: +1.734.677.1774 5


6<br />

Detectx ®<br />

Acetylcholinesterase (AChe) Fluorescent Activity <strong>kit</strong><br />

Catalog No: K015-F1 (2 Plate)<br />

FeAtures<br />

f Use Measure AChE Activity in 20 Minutes<br />

f<br />

f<br />

f<br />

Sample CSF, Serum, Plasma, and solublized RBC ghosts<br />

Species Human and other mammalian species<br />

Samples/Kit 90 in Duplicate<br />

iNtroDuCtioN<br />

Acetylcholinesterases (AChE) appear critical to both development and function of the nervous system. The use of AChE<br />

inhibitors as therapeutic agents and pesticides has spurred detailed investigations of cholinesterases. Acetylcholine (ACh) is<br />

an essential neurotransmitter in the central and peripheral nervous systems. In the brain multiple areas exist where cholinergic<br />

neurons are concentrated. Nicotinic and muscarinic acetylcholine receptors are recognized as binding and effector proteins<br />

to mediate chemical neurotransmission at neurons, ganglia, heart and smooth muscle fibers and glands.<br />

The DetectX® Acetylcholinesterase Activity <strong>kit</strong> is designed to quantitatively measure AChE activity in a variety of samples such<br />

as diluted CSF, serum, plasma and RBC ghosts from a number of species. A human AChE standard is provided to generate<br />

a standard curve for the assay. The <strong>kit</strong> utilizes a proprietary non-fluorescent molecule, ThioStar®, that covalently binds to<br />

the thiol product of the reaction between the AChE Substrate and AChE in the standards or samples, yielding a fluorescent<br />

product read at 510 nm in a fluorescent plate reader with excitation at 390 nm. The readout of this AChE activity assay is<br />

purely chemical and no other enzymes are involved, and therefore few interferents will affect the readings obtained.<br />

typiCAL DAtA<br />

reLAteD proDuCts<br />

Butyrylcholinesterase Activity Kit Catalog No. K016-F1 Page 8<br />

Hemoglobin Dual Range Detection Kit Catalog No. K013-H1 Page 25<br />

Urea Nitrogen (BUN) Detection Kit Catalog No. K024-H1 Page 41<br />

ThioStar ® Detection System Catalog No. L002 Page 42<br />

www.ArborAssays.com | info@ArborAssays.com | Ph: +1.734.677.1774


Detectx ®<br />

Atrial Natriuretic peptide (ANp) <strong>immunoassay</strong> (<strong>eiA</strong>) <strong>kit</strong>s<br />

FeAtures<br />

f Use Quantitate ANP in a variety of samples<br />

f<br />

f<br />

f<br />

f<br />

Sample Plasma or Urine samples<br />

Samples/<strong>kit</strong> 40 in Duplicate in 1 plate <strong>kit</strong>, 232 with 5 plate <strong>kit</strong><br />

Range 180-0.74 ng/mL<br />

Stability Liquid, 4°C Stable Reagents<br />

Catalog No: K026-H1 (1 Plate) K026-H5 (5 Plate)<br />

iNtroDuCtioN<br />

Atrial natriuretic peptide (ANP), a peptide hormone secreted mostly by cardiac myocytes, is a potent natriuretic, diuretic<br />

and vasodilatory peptide that contributes to blood pressure and volume homeostasis. ANP is released by myocytes in<br />

response to atrial distension. Upon binding to cell surface receptors (NPR-A, B, and C, also termed guanylyl cyclase-A and B<br />

receptors), ANP acts through generation of cyclic guanosine monophosphate (cGMP). Atrial natriuretic peptide demonstrates<br />

hemodynamic and glomerular effects, which increase sodium and water load delivery to the tubules, and the inhibition of the<br />

release of renin, aldosterone and vasopressin.<br />

The DetectX® Atrial Natriuretic Peptide (ANP) Immunoassay <strong>kit</strong>s are designed to measure ANP present in plasma and<br />

urine samples. An ANP standard is provided for the assays. Standards or samples are pipetted into a coated microtiter<br />

plate. An ANP conjugate is added to the wells. The binding reaction is initiated by the addition of a rabbit polyclonal<br />

antibody to ANP. After an hour incubation, the plate is washed and substrate is added. The substrate reacts with the bound<br />

ANP conjugate. After a short incubation the intensity of the generated color is measured at 450 nm.<br />

typiCAL DAtA<br />

reLAteD proDuCts<br />

Cyclic AMP Direct EIA and CLIA Kits Catalog No. K019-H/C Page 15<br />

Cyclic GMP Direct EIA and CLIA Kits Catalog No. K020-H/C Page 16<br />

Phosphokinase A (PKA) Activity Kit Catalog No. K027-H1 Page 35<br />

Nitric Oxide Detection Kit Catalog No. K023-H1 Page 28<br />

www.ArborAssays.com | info@ArborAssays.com | Ph: +1.734.677.1774 7


8<br />

Detectx ®<br />

butyrylcholinesterase (bChe) Fluorescent Activity <strong>kit</strong><br />

Catalog No: K016-F1 (2 Plate)<br />

FeAtures<br />

f Use Measure BChE Activity in 20 Minutes<br />

f<br />

f<br />

f<br />

Sample CSF, Serum, and Plasma<br />

Species Human and other mammalian species<br />

Samples/Kit 88 in Duplicate<br />

iNtroDuCtioN<br />

Butyrylcholinesterase (BChE) belongs to the same structural class of proteins as acetylcholinesterase (AChE). The 440kDa<br />

tetrameric glycoprotein is predominantly found in blood, kidneys, intestine, liver, lung, heart and the central nervous system.<br />

BChE preferentially acts on butyrylcholine, but also hydrolyzes acetylcholine. BChE hydrolyzes ester-containing drugs<br />

and scavenges cholinesterase inhibitors, such as succinylcholine, before they have a chance to reach synaptic targets. A<br />

deficiency of BChE can result in delayed metabolism of various drugs, such as cocaine, and treatment with doses of BChE<br />

can help in overcoming their physiological effects.<br />

In Alzheimer’s disease the reduction in choline acetyltransferase leads to a decrease in acetylcholine and acetylcholinesterase<br />

activity, which appears to cause an increase in BChE activity. Selective BChE inhibitors prevent the formation of new betaamyloid<br />

plaques, which are created by BChE cleaving APP to beta-amyloid. BChE-positive neurons project to the frontal<br />

cortex portion of the brain. BChE may have roles in attention, executive function, emotional memory and behaviour. As<br />

dementia advances, BChE activity has been shown to increase, while AChE activity decreases, leaving the potential for<br />

BChE activity to be used as a biomarker for progression or target for future therapies.<br />

The DetectX® Butyrylcholinesterase Activity <strong>kit</strong> is designed to quantitatively measure BChE activity in a variety of samples<br />

such as diluted CSF, serum, and plasma from a number of species. A human BChE standard is provided to generate a<br />

standard curve for the assay. The <strong>kit</strong> utilizes a proprietary non-fluorescent molecule, ThioStar®, that covalently binds to the<br />

thiol product of the reaction between the BChE Substrate and BChE in the standards or samples, yielding a fluorescent<br />

product read at 510 nm in a fluorescent plate reader with excitation at 390 nm. The readout of this BChE activity assay is<br />

purely chemical and no other enzymes are involved, and few interferents will affect the readings obtained.<br />

typiCAL DAtA<br />

reLAteD proDuCts<br />

Acetylcholinesterase Activity Kit Catalog No. K015-F1 Page 6<br />

Hemoglobin Dual Range Detection Kit Catalog No. K013-H1 Page 25<br />

Urea Nitrogen (BUN) Detection Kit Catalog No. K024-H1 Page 41<br />

ThioStar® Detection System Catalog No. L002 Page 42<br />

www.ArborAssays.com | info@ArborAssays.com | Ph: +1.734.677.1774


FeAtures<br />

f Complete Everything needed to measure Catalase activity<br />

f<br />

f<br />

f<br />

Stable Liquid, 4°C stable reagents<br />

Rapid 45 minute assay<br />

Economical 89 Samples<br />

Detectx ®<br />

Catalase Fluorescent Activity <strong>kit</strong><br />

Catalog No: K033-F1 (2 Plate)<br />

iNtroDuCtioN<br />

Hydrogen peroxide, H 2 O 2 is one of the most frequently occurring reactive oxygen species. It is formed either in the environment<br />

or as a by-product of aerobic metabolism, superoxide formation and dismutation, or as a product of oxidase activity. Both<br />

excessive hydrogen peroxide and its decomposition product hydroxyl radical, formed in a Fenton-type reaction, are harmful<br />

for most cell components. One of the most efficient ways of removing peroxide is through the enzyme catalase, which is<br />

encoded by a single gene, and is highly conserved among species. Mammals, including humans and mice, express catalase<br />

in all tissues, and a high concentration of catalase can be found in the liver, kidneys and erythrocytes. The expression is<br />

regulated at transcription, post-transcription and post-translation levels. High catalase activity is detected in peroxisomes.<br />

More recently, short wavelength UV radiation has been shown to produce reactive oxygen species (ROS) through the action<br />

of catalase. This response is thought to act as a mechanism to protect DNA by converting damaging UV radiation into ROS<br />

species that can be metabolized and detoxified by cellular antioxidant enzymes.<br />

The DetectX® Catalase Activity Kit is designed to quantitatively measure catalase activity in a variety of samples. A<br />

bovine catalase standard is provided. Samples are diluted in the provided Assay Buffer and added to the wells of a half<br />

area black plate. Hydrogen peroxide is added to each well and the plate incubated at room temperature for 30 minutes.<br />

The supplied Fluorescent Detection Reagent is added, followed by diluted horseradish peroxidase and incubated at<br />

room temperature for 15 minutes. The HRP reacts with the substrate in the presence of hydrogen peroxide to produce a<br />

fluorescent product. The fluorescent product is read at 590 nm with excitation at 570 nm. Increasing levels of catalase<br />

in the samples causes a decrease in H 2 O 2 concentration and a reduction in fluorescent product.<br />

typiCAL DAtA<br />

Mean FLU<br />

40,000<br />

35,000<br />

30,000<br />

25,000<br />

20,000<br />

Mean FLU<br />

15,000<br />

10,000<br />

5,000<br />

0<br />

0 1.0 2.0 3.0 4.0 5.0<br />

Catalase Activity (U/mL)<br />

reLAteD proDuCts<br />

Hydrogen Peroxide Fluorescent Detection Kit Catalog No. K034-F1 Page 27<br />

Glutathione Fluorescent Detection Kits Catalog No. K006-F1/F5 Page 22<br />

Superoxide Dismutase Activity Kit Catalog No. K028-H1 Page 38<br />

Glutathione Colorimetric Detection Kit Catalog No. K006 Page 22<br />

www.ArborAssays.com | info@ArborAssays.com | Ph: +1.734.677.1774 9


10<br />

Detectx ®<br />

Ceruloplasmin (Cp) Colorimetric Activity <strong>kit</strong><br />

Catalog No: K035-H1 (2 Plate) Patents Pending<br />

FeAtures<br />

f Use Non-invasive Multi-Species Pregnancy Marker<br />

f<br />

f<br />

f<br />

Sample Urine<br />

Species Human and other mammalian species<br />

Samples/Kit 88 in Duplicate with Full Standard Curve<br />

iNtroDuCtioN<br />

Ceruloplasmin (Cp) is a multicopper oxidase enzyme involved in the safe handling of oxygen in some metabolic pathways<br />

of vertebrates. Discovered in 1948, a blue protein from the a2-globulin fraction of human serum possessing oxidase activity<br />

towards aromatic diamines and catechol was purified by Holmberg and Laurell. It was denoted ceruloplasmin, literally<br />

meaning ‘a blue substance from plasma’. Specialized copper sites have been recruited during evolution to provide longrange<br />

electron transfer reactivity and oxygen binding and activation in proteins destined to cope with oxygen reactivity in<br />

different organisms. Ceruloplasmin belongs to the family of multicopper oxidases which are among the few enzymes able<br />

to bind molecular oxygen to perform its complete reduction to water. Ceruloplasmin contains 95% of the copper in serum.<br />

Cp found in serum is expressed in the liver, but it is also expressed in the brain, lung, spleen and testis.<br />

Aceruloplasminaemia is an autosomal recessive disorder of iron metabolism characterized by the complete absence of<br />

ceruloplasmin. The role of Cp in tissue iron overload and the subsequent clinical findings of diabetes, retinal degeneration<br />

and neurodegeneration has been associated with iron overload in aceruloplasminaemic patients. Thus it is clearly indicated<br />

that ceruloplasmin plays an essential role in iron metabolism. Ceruloplasmin is also associated with reproduction. Copperdeficient<br />

female rats seem to be protected against mortality. This protection has been suggested to be provided by<br />

estrogens, since estrogens alter the subcellular distribution of copper in the liver, an increase in plasma copper levels and<br />

subsequent ceruloplasmin synthesis.<br />

The DetectX® Ceruloplasmin Activity Kit is designed to quantitatively measure ceruloplasmin activity in urine samples. A<br />

human ceruloplasmin standard is provided. Samples are diluted in the provided Assay Buffer and added to the wells of a<br />

half area clear plate. The reconstituted Ceruloplasmin Substrate is added and the plate is incubated at 30°C for 60 minutes.<br />

The ceruloplasmin in the standards and samples react with the substrate to produce a colored product. The optical density<br />

is read at 560 nm. Increasing levels of ceruloplasmin in the samples cause an increase in the fuschia (pink/purple) product.<br />

The results are expressed in terms of units of ceruloplasmin activity per mL.<br />

typiCAL DAtA<br />

reLAteD proDuCts<br />

Estradiol EIA Kits Catalog No. K030-H1/H5 Page 18<br />

PGFM EIA Kits Catalog No. K022-H1/H5 Page 32<br />

Estrone-3-Glucuronide (E1G) EIA Kits Catalog No. K036-H1/H5 Page 20<br />

Pregnanediol Glucuronide (PDG) EIA Kits Catalog No. K037-H1/H5 Page 33<br />

www.ArborAssays.com | info@ArborAssays.com | Ph: +1.734.677.1774


Detectx ®<br />

Corticosterone <strong>immunoassay</strong> (<strong>eiA</strong> and CLiA) <strong>kit</strong>s<br />

Colorimetric Catalog No: K014-H1 (1 Plate) K014-H5 (5 Plate)<br />

Chemiluminescent Catalog No: K014-C1 (1 Plate) K014-C5 (5 Plate)<br />

FeAtures<br />

f Use Non-Invasive Fecal Extracts, Serum, Plasma and TCM<br />

f Sample Size 2 µL serum or plasma needed<br />

f<br />

f<br />

Fecal/Urine Mice, rats, apes, cattle, deer, equids, felids, and ungulates<br />

Samples/Kit 39 in Duplicate in 1 plate <strong>kit</strong>, 231 with 5 plate <strong>kit</strong><br />

iNtroDuCtioN<br />

Corticosterone (C 21 H 30 O 4 , Kendall’s Compound ‘B’) is a glucocorticoid secreted by the cortex of the adrenal gland.<br />

Corticosterone is produced in response to stimulation of the adrenal cortex by ACTH and is the precursor of aldosterone.<br />

Corticosterone is a major indicator of stress and is the major stress steroid produced in non-human mammals. Studies<br />

involving corticosterone and levels of stress include impairment of long term memory retrieval, chronic corticosterone<br />

elevation due to dietary restrictions and in response to burn injuries. In addition to stress levels, corticosterone is believed<br />

to play a decisive role in sleep-wake patterns.<br />

Corticosterone<br />

The DetectX® Corticosterone Immunoassay <strong>kit</strong>s is designed to quantitatively measure corticosterone present in extracted<br />

dried fecal samples, serum, plasma and tissue culture media samples. This <strong>kit</strong> measures total corticosterone in serum,<br />

plasma and in extracted fecal samples. A corticosterone standard is provided for the assay. Standards or samples are<br />

pipetted into a coated microtiter plate. A corticosterone-peroxidase conjugate is added to the wells. The binding reaction<br />

is initiated by the addition of a sheep polyclonal antibody to corticosterone. After an hour incubation the plate is washed<br />

and substrate is added. The substrate reacts with the bound corticosterone-peroxidase conjugate. For the EIA <strong>kit</strong>s, the<br />

reaction is stopped after a short incubation the plate read at 450 nm. For the CLIA <strong>kit</strong>s, the light is read immediately.<br />

typiCAL DAtA<br />

reLAteD proDuCts<br />

Cortisol EIA Kits Catalog No. K003-H1/H5 Page 12<br />

Cortisone CLIA Kits Catalog No. K017-C1/C5 Page 13<br />

Urinary Creatinine Kits Catalog No. K002-H1/H5 Page 14<br />

Hemoglobin Dual Range Kit Catalog No. K013-H1 Page 25<br />

Urea Nitrogen (BUN) Kit Catalog No. K024-H1 Page 41<br />

www.ArborAssays.com | info@ArborAssays.com | Ph: +1.734.677.1774 11


12<br />

Detectx ®<br />

Cortisol <strong>immunoassay</strong> (<strong>eiA</strong>) <strong>kit</strong>s<br />

Strip Wells Catalog No.: K003-H1 (1 Plate) K003-H5 (5 Plate)<br />

Whole Plate Catalog No.: K003-H1W (1 Plate) K003-H5W (5 Plate)<br />

FeAtures<br />

f Use Non-Invasive Urine or Fecal Extracts, Serum, Plasma, Saliva and TCM<br />

f<br />

f<br />

f<br />

f<br />

Serum/Plasma Tested in human through fish samples<br />

Fecal Samples Apes, cattle, deer, equids, felids, and ungulates<br />

Samples/Kit 39 in Duplicate in 1 plate <strong>kit</strong>, 231 with 5 plate <strong>kit</strong><br />

Stability All Liquid Reagents Stable at 4°C<br />

iNtroDuCtioN<br />

Cortisol, C 21 H 30 O 5 , (hydrocortisone, compound F) is the primary glucocorticoid produced and secreted by the adrenal<br />

cortex. It is often referred to as the “stress hormone” as it is involved in the response to stress and it affects blood<br />

pressure, blood sugar levels, and other actions of stress adaptation. Immunologically, cortisol functions as an important<br />

anti-inflammatory and plays a role in hypersensitivity, immunosuppression, and disease resistance. In the metabolic aspect,<br />

cortisol promotes gluconeogenesis, liver glycogen deposition, and the reduction of glucose utilization. Production of<br />

cortisol follows an ACTH-dependent circadian rhythm, with a peak level in the morning and decreasing levels throughout<br />

the day. Most serum cortisol, all but about 4%, is bound to proteins including corticosteroid binding globulin and serum<br />

albumin. Only free cortisol is available to most receptors and it is through these receptors that physiological processes<br />

are modulated. Abnormal cortisol levels are being evaluated for correlation with a variety of different conditions, such<br />

as prostate cancer, depression, and schizophrenia. It is already known that abnormal levels of cortisol are involved in<br />

Cushing’s Syndrome and Addision’s disease.<br />

Cortisol<br />

The DetectX® Cortisol Immunoassay <strong>kit</strong> measures cortisol present in extracted fecal samples, urine, serum, plasma, saliva<br />

and TCM samples. This <strong>kit</strong> measures total cortisol in serum and plasma and in extracted fecal samples. A cortisol standard,<br />

calibrated to the NIST cortisol standard, is provided for the assay. Standards or samples are pipetted into a coated clear<br />

microtiter plate. A cortisol-peroxidase conjugate is added to the wells. The binding reaction is initiated by the addition of a<br />

monoclonal antibody to cortisol. After an hour incubation the plate is washed and substrate is added. The substrate reacts<br />

with the bound cortisol-peroxidase conjugate and the color is measured at 450 nm.<br />

typiCAL DAtA<br />

reLAteD proDuCts<br />

Corticosterone EIA Kits Catalog No. K014-H1/H5 Page 11<br />

Cortisone CLIA Kit Catalog No. K017-C1/C5 Page 13<br />

Urinary Creatinine Detection Kit Catalog No. K002-H1/H5 Page 14<br />

Hemoglobin Dual Range Detection Kit Catalog No. K013-H1 Page 25<br />

www.ArborAssays.com | info@ArborAssays.com | Ph: +1.734.677.1774


Detectx ®<br />

Cortisone Chemiluminescent <strong>immunoassay</strong> (CLiA) <strong>kit</strong>s<br />

FeAtures<br />

f Use Measure the Activity of 11ß-HSD Enzymes<br />

f<br />

f<br />

f<br />

Samples Serum, Saliva, Urine and Fecal Extracts<br />

Samples/Kit 37 in Duplicate in 1 plate <strong>kit</strong>, 229 with 5 plate <strong>kit</strong><br />

Stability All Liquid Reagents Stable at 4°C<br />

Catalog No: K017-C1 (1 Plate) K017-C5 (5 Plate)<br />

iNtroDuCtioN<br />

Cortisone (C 21 H 28 O 5 , Kendall’s Compound ‘E’) was identified by extraction from bovine suprarenal gland tissue. Compound<br />

E was soon identified as cortisone. The more active Compound F, cortisol, and cortisone vary due to the activity of<br />

two 11ß-hydroxysteroid dehydrogenases (11ß-HSD). 11ß-HSD1 is found primarily in the liver where it converts cortisone<br />

to cortisol while 11ß-HSD2 is found in tissues such as the kidney where cortisol receptor binding is required. 11ß-HSD2<br />

deactivates cortisol to cortisone, prohibiting receptor activation. This glucocorticoid “shuttle” helps to initiate and regulate<br />

the anti-inflammatory response. Monitoring the ratio of cortisone:cortisol has applications in diabetes, obesity, metabolic<br />

syndrome, osteoporosis, and chronic fatigue syndrome in addition to adrenal diseases.<br />

Cortisone<br />

The DetectX® Cortisone Immunoassay <strong>kit</strong> measures cortisone present in serum, saliva, urine and extracted dried fecal<br />

samples. This <strong>kit</strong> measures total cortisone in serum, urine and in extracted fecal samples. A cortisone standard is provided.<br />

Standards or samples are pipetted into a coated white microtiter plate. A cortisone-peroxidase conjugate is added to the<br />

wells. The binding reaction is initiated by the addition of a polyclonal antibody to cortisone. After a 2 hour incubation the<br />

plate is washed and substrate is added. The substrate reacts with the bound cortisone-peroxidase conjugate. The emitted<br />

light is read immediately for 0.1 sec per well in a multi-label plate reader.<br />

typiCAL DAtA<br />

Cortisone<br />

reLAteD proDuCts<br />

Cortisol EIA Kits Catalog No. K003-H1/H5 Page 12<br />

Corticosterone EIA Kits Catalog No. K014-H1/H5 Page 11<br />

Urinary Creatinine Detection Kit Catalog No. K002-H1/H5 Page 14<br />

Hemoglobin Dual Range Detection Kit Catalog No. K013-H1 Page 25<br />

Urea Nitrogen (BUN) Detection Kit Catalog No. K024-H1 Page 41<br />

www.ArborAssays.com | info@ArborAssays.com | Ph: +1.734.677.1774 13


14<br />

Detectx ®<br />

Creatinine urine and serum Detection <strong>kit</strong>s<br />

Urinary Creatinine Catalog No.: K002-H1 (2 Plate) K002-H5 (10 Plate)<br />

Serum Creatinine Catalog No.: KB02-H1/H2 (2/4 Plate) KB02-H1D (384 well)<br />

FeAtures urinary <strong>kit</strong> serum <strong>kit</strong><br />

f Use Urine Volume Kidney Damage<br />

f<br />

f<br />

f<br />

f<br />

Species Species Independent Most species<br />

Calibrated NIST Standard Reference #914a NIST Standard Reference #914a<br />

Samples/Kit 88 or 472 in Duplicate 91/187 in Duplicate, 187 for Low Volume 384 well<br />

Stability All Liquid Reagents Stable at 4°C All Liquid Reagents Stable at 4°C<br />

iNtroDuCtioN<br />

Creatinine (2-amino-1-methyl-5H-imadazol-4-one) is a metabolite of phosphocreatine (p-creatine), a molecule used as a<br />

store for high-energy phosphate that can be utilized by tissues for the production of ATP. Creatine and p-creatine are<br />

converted non-enzymatically to the metabolite creatinine, which diffuses into the blood and is excreted by the kidneys. Its<br />

formation occurs at a rate that is relatively constant and, as intra-individual variation is


Detectx ®<br />

Cyclic Amp Direct <strong>immunoassay</strong> (<strong>eiA</strong> and CLiA) <strong>kit</strong>s<br />

Colorimetric Catalog No: K019-H1 (1 Plate) K019-H5 (5 Plate)<br />

Chemiluminescent Catalog No: K019-C1 (1 Plate) K019-C5 (5 Plate)<br />

FeAtures<br />

f Use Measure cAMP in cells, saliva, urine, plasma and tissue<br />

f<br />

f<br />

f<br />

f<br />

Convenient Lyse, stabilize and measure in one step - Results in < 3 Hours<br />

Sensitive EIA <strong>kit</strong> : 4.2 fmol cAMP CLIA: 0.75 fmol cAMP<br />

Samples/Kit EIA <strong>kit</strong>: 39 or 231 in Duplicate CLIA: 38 or 230 in Duplicate<br />

Stability All Liquid Reagents Stable at 4°C<br />

iNtroDuCtioN<br />

Adenosine-3’,5’-cyclic monophosphate, or cyclic AMP (cAMP), C 10 H 12 N 5 O 6 P, is one of the most important second messengers<br />

and a key intracellular regulator. Discovered by Sutherland and Rall in 1957, it functions as a mediator of activity for a number<br />

of hormones, including epinephrine, glucagon, and ACTH. Adenylate cyclase is activated by the hormones glucagon and<br />

adrenaline and by G protein. Liver adenylate cyclase responds more strongly to glucagon, and muscle adenylate cyclase<br />

responds more strongly to adrenaline. cAMP decomposition into AMP is catalyzed by the enzyme phosphodiesterase. In<br />

the Human Metabolome Database there are 166 metabolic enzymes listed that convert cAMP. Other biological actions of<br />

cAMP include regulation of innate immune functioning, axon regeneration, cancer, and inflammation.<br />

Cyclic Amp<br />

The DetectX® Direct Cyclic AMP (cAMP) Immunoassay <strong>kit</strong>s are designed to quantitatively measure cAMP present in lysed<br />

cells or tissue, EDTA and heparin plasma, urine, saliva and tissue culture media samples. The <strong>kit</strong> is unique in that all<br />

samples are diluted into an acidic Sample Diluent for cAMP measurement. The treated samples are stable and endogenous<br />

phosphodiesterases are inactivated in the Sample Diluent. A microtiter plate coated with an antibody to capture IgG is<br />

provided. Samples are pipetted into the primed wells. A cAMP-peroxidase conjugate is added and the binding reaction<br />

is initiated by the addition of antibody to cAMP. After a 2 hour incubation, the plate is washed and substrate is added.<br />

The substrate reacts with the bound cAMP-peroxidase conjugate. For the EIA <strong>kit</strong>s, the reaction is stopped after a short<br />

incubation the plate read at 450 nm. For the CLIA <strong>kit</strong>s, the light is read immediately.<br />

typiCAL DAtA<br />

reLAteD proDuCts<br />

Cyclic GMP Direct EIA and CLIA Kits Catalog No. K020-H/C Page 16<br />

Phosphokinase A (PKA) Activity Kit Catalog No. K027-H1 Page 35<br />

IBMX - Pan-specific PDE Inhibitor Catalog No. P019-100MG/1G Page 44/45<br />

www.ArborAssays.com | info@ArborAssays.com | Ph: +1.734.677.1774 15


16<br />

Detectx ®<br />

Cyclic gmp Direct <strong>immunoassay</strong> (<strong>eiA</strong> and CLiA) <strong>kit</strong>s<br />

Colorimetric Catalog No:. K020-H1 (1 Plate) K020-H5 (5 Plate)<br />

Chemiluminescent Catalog No.: K020-C1 (1 Plate) K020-C5 (5 Plate)<br />

FeAtures<br />

f Use Measure cGMP in cells, saliva, urine, plasma and tissue<br />

f<br />

f<br />

f<br />

f<br />

Convenient Lyse, stabilize and measure in one step<br />

Sensitive EIA <strong>kit</strong> : 9.4 fmol cGMP CLIA: 1.15 fmol cGMP<br />

Samples/Kit EIA <strong>kit</strong>: 39 or 231 in Duplicate CLIA: 39 or 231 in Duplicate<br />

Stability All Liquid Reagents Stable at 4°C<br />

iNtroDuCtioN<br />

Guanosine 3’, 5’-cyclic monophosphate (cyclic GMP; cGMP) is a critical and multifunctional second messenger present at<br />

levels typically 10-100 fold lower than cAMP in most tissues. Intracellular cGMP is formed by the action of the enzyme<br />

guanylate cyclase on GTP and degraded through phosphodiesterase hydrolysis. Guanylate cyclases (GC) are either soluble<br />

or membrane bound. Soluble GCs are nitric oxide responsive, whereas membrane bound GCs respond to hormones such as<br />

acetylcholine, insulin and oxytocin. Other chemicals like serotonin and histamine also cause an increase in cGMP levels. Cyclic<br />

GMP regulates cellular composition through cGMP-dependent kinase, cGMP-dependent ion channels or transporters, and<br />

through its hydrolytic degradation by phosphodiesterase.<br />

Cyclic gmp<br />

The DetectX® Direct Cyclic GMP (cGMP) Immunoassay <strong>kit</strong>s are designed to quantitatively measure cGMP present in lysed cells or<br />

tissue, EDTA and heparin plasma, urine, saliva and tissue culture media samples. The <strong>kit</strong> is unique in that all samples are diluted<br />

into an acidic Sample Diluent for cGMP measurement. The cGMP in the samples is stable and endogenous phosphodiesterases are<br />

inactivated in the Sample Diluent. A cGMP standard is provided. A microtiter plate coated with an antibody to capture mouse IgG<br />

is provided and a neutralizing Plate Primer solution is added to all wells. Samples, either with or without acetylation, are pipetted<br />

into the primed wells. A cGMP-peroxidase conjugate is added and the binding reaction is initiated by the addition of a mouse<br />

monoclonal antibody to cGMP. After a 2 hour incubation for the EIA or a 16 hour for the CLIA, the plate is washed and substrate<br />

is added. The substrate reacts with the bound cGMP-peroxidase conjugate. For the EIA <strong>kit</strong>s, the reaction is stopped after a short<br />

incubation the plate read at 450 nm. For the CLIA <strong>kit</strong>s, the light emitted is read immediately.<br />

typiCAL DAtA<br />

%B/B0<br />

100<br />

90<br />

80<br />

70<br />

60<br />

50<br />

%B/B0<br />

40<br />

30<br />

20<br />

10<br />

0<br />

0.01 0.1 1 10 100<br />

Cyclic GMP Conc. (pmol/mL)<br />

H1/H5<br />

H1/H5 Acetylated<br />

C1/C5<br />

C1/C5 Acetylated<br />

reLAteD proDuCts<br />

ANP EIA Kits Catalog No. K026-H1 Page 7<br />

Cyclic AMP Direct EIA and CLIA Kits Catalog No. K019-H/C Page 15<br />

Phosphokinase A (PKA) Activity Kit Catalog No. K027-H1 Page 35<br />

IBMX - Pan-specific PDE Inhibitor Catalog No. P019-100MG/1G Page 44/45<br />

www.ArborAssays.com | info@ArborAssays.com | Ph: +1.734.677.1774


FeAtures<br />

f Use Kidney Injury Marker<br />

f<br />

f<br />

f<br />

f<br />

Sample Type Serum, Plasma, Urine or TCM<br />

Samples/Kit 40 Samples in Duplicate<br />

Detectx ®<br />

human Cystatin C enzyme <strong>immunoassay</strong> (<strong>eiA</strong>) <strong>kit</strong><br />

Stability All Liquid Reagents Stable at 4°C<br />

Performance 0.156-10 ng/mL in 2 Hours<br />

Catalog No.: K012-H1 (1 Plate)<br />

iNtroDuCtioN<br />

Cystatin C is a non-glycosylated protein of low molecular weight (13kDa) in the cystatin superfamily. Cystatin C is produced<br />

at a constant rate in all nucleated cells, secreted from cells and thus found in most body fluids. Cystatin C belongs to the<br />

cysteine proteinase inhibitor group and is associated with several pathological conditions. Imbalance between Cystatin C<br />

and cysteine proteinases is associated with diseases such as inflammation, renal failure, cancer, Alzheimer’s disease, multiple<br />

sclerosis and hereditary Cystatin C amyloid angiopathy. Cystatin C is removed from blood plasma by glomerular filtration in<br />

the kidneys. It is reabsorbed by the proximal tubular cells and degraded. There is a linear relationship between the reciprocal<br />

Cystatin C concentration in plasma and the glomerular filtration rate (GFR). Cystatin C is suggested to be a better marker for<br />

GFR than serum creatinine as its serum concentration is not affected by factors such as age, gender and body mass. There is<br />

association of Cystatin C levels with the incidence of myocardial infarction, coronary death and angina pectoris, presenting a<br />

risk factor for secondary cardiovascular events.<br />

The DetectX® Human Cystatin C Immunoassay <strong>kit</strong> is designed to quantitatively measure human Cystatin C present in biological<br />

samples and tissue culture media. A human Cystatin C standard is used to generate a standard curve for the assay. Standards<br />

or diluted samples are pipetted into a clear microtiter plate coated with a monoclonal antibody to capture the Cystatin C.<br />

After a 60 minute incubation, the plate is washed and a peroxidase conjugated Cystatin C monoclonal antibody is added. The<br />

plate is incubated for 30 minutes and washed. Substrate is then added to the plate, which reacts with the bound Cystatin C<br />

conjugate. The color reaction is stopped and the intensity read at 450 nm.<br />

typiCAL DAtA<br />

Optical Density<br />

1.75<br />

1.50<br />

1.25<br />

1.00<br />

0.75<br />

Optical Density<br />

0.50<br />

0.25<br />

0.00<br />

0 1 2 3 4 5 6 7 8 9 10<br />

Cystatin C Conc. (ng/mL)<br />

reLAteD proDuCts<br />

Serum Creatinine Detection Kits Catalog No. KB02-H1/H2 Page 14<br />

Urinary Retinol Binding Protein EIA Kit Catalog No. KU04-H1 Page 37<br />

Hemoglobin Dual Range Detection Kit Catalog No. K013-H1 Page 25<br />

Urea Nitrogen (BUN) Detection Kit Catalog No. K024-H1 Page 41<br />

www.ArborAssays.com | info@ArborAssays.com | Ph: +1.734.677.1774 17


18<br />

Detectx ®<br />

estradiol <strong>immunoassay</strong> (<strong>eiA</strong>) <strong>kit</strong>s<br />

Catalog No.: K030-H1 (1 Plate) K030-H5 (5 Plate)<br />

FeAtures<br />

f Use Non-Invasive Fecal Extracts, and TCM<br />

f<br />

f<br />

f<br />

Serum/Plasma Tested in human through fish samples<br />

Fecal Samples Apes, cattle, deer, equids, felids, and ungulates<br />

Samples/Kit 41 in Duplicate in 1 plate <strong>kit</strong>, 233 with 5 plate <strong>kit</strong><br />

iNtroDuCtioN<br />

Estradiol (E2 or 17ß-estradiol, also oestradiol) is the predominant sex hormone present in females. It is also present in<br />

males, being produced as an active metabolic product of testosterone. It represents the major estrogen in humans.<br />

Estradiol has not only a critical impact on reproductive and sexual functioning, but also affects other organs. Serum<br />

estradiol measurement in women reflects primarily the activity of the ovaries. As such, they are useful in the detection of<br />

baseline estrogen in women with amenorrhea or menstrual dysfunction and to detect the state of hypoestrogenicity and<br />

menopause. Furthermore, estrogen monitoring during fertility therapy assesses follicular growth and is useful in monitoring<br />

the treatment. Estrogen-producing tumors will demonstrate persistent high levels of estradiol and other estrogens. In<br />

precocious puberty, estradiol levels are inappropriately increased.<br />

17ß-estradiol<br />

The DetectX® Estradiol Immunoassay <strong>kit</strong> is designed to quantitatively measure estradiol present in extracted dried fecal<br />

samples and tissue culture media samples. This <strong>kit</strong> measures total estradiol in extracted serum and plasma and in extracted<br />

fecal samples. An estradiol standard is provided for the assay. Standards or samples are pipetted into a microtiter plate.<br />

A estradiol-peroxidase conjugate is added to the wells. The binding reaction is initiated by the addition of a polyclonal<br />

antibody. After a 2 hour incubation the plate is washed and substrate is added. The substrate reacts with the bound<br />

estradiol-peroxidase conjugate. After a short incubation the intensity of the color is measured at 450 nm.<br />

typiCAL DAtA<br />

reLAteD proDuCts<br />

Estrone EIA Kits Catalog No. K031-H1/H5 Page 19<br />

Estrone-3- Glucuronide (E1G)EIA Kits Catalog No. K036-H1/H5 Page 20<br />

Progesterone EIA Kits Catalog No. K025-H1/H5 Page 34<br />

Urinary Creatinine Detection Kit Catalog No. K002-H1/H5 Page 14<br />

Hemoglobin Dual Range Detection Kit Catalog No. K013-H1 Page 25<br />

Urea Nitrogen (BUN) Detection Kit Catalog No. K024-H1 Page 41<br />

www.ArborAssays.com | info@ArborAssays.com | Ph: +1.734.677.1774


FeAtures<br />

f Non-Invasive Measures Estrone, 3-sulfate and 3-glucuronide<br />

f<br />

f<br />

f<br />

Adaptable Tested in a Wide Variety of Species<br />

Samples/Kit 39 in Duplicate in 1 plate <strong>kit</strong>, 231 with 5 plate <strong>kit</strong><br />

Stability All Liquid Reagents Stable at 4°C<br />

Detectx ®<br />

estrone enzyme <strong>immunoassay</strong> (<strong>eiA</strong>) <strong>kit</strong>s <strong>kit</strong>s<br />

Catalog No.: K031-H1 (1 Plate) K031-H5 (5 Plate)<br />

iNtroDuCtioN<br />

Estrone, C 18 H 22 O 2 , also known as E1 or osterone (3-hydroxy-1,3,5(10)-estratrien-17-one) is a C-18 steroid hormone. Estrone<br />

is one of the three naturally occurring estrogens, the others being estradiol and estriol. Estrone is produced primarily from<br />

androstenedione originating from the gonads or the adrenal cortex and from estradiol by 17-hydroxysteroid dehydrogenase.<br />

Estrone concentrations in premenopausal mammals fluctuate according to the menstrual cycle. In premenopausal women,<br />

more than 50% of the estrone is secreted by the ovaries. Interconversion of estrone and estradiol also occurs in peripheral<br />

tissue. In humans, during the follicular phase of the menstrual cycle estrone levels increase slightly.<br />

estrone<br />

The DetectX® Estrone Immunoassay <strong>kit</strong> is designed to quantitatively measure estrone present in extracted dried fecal<br />

samples, urine and tissue culture media samples. An estrone standard is provided to generate a standard curve for the<br />

assay. Standards or diluted samples are pipetted into an antibody coated clear microtiter plate. An estrone-peroxidase<br />

conjugate is added to the standards and samples. The binding reaction is initiated by the addition of a polyclonal antibody<br />

to estrone. After a 2 hour incubation the plate is washed and substrate is added. The substrate reacts with the bound<br />

estrone-peroxidase conjugate. After a short incubation, the reaction is stopped and the color read at 450nm. The <strong>kit</strong><br />

is unique as it measures both non-conjugated estrone, estrone-3-sulfate and estrone 3-glucuronide in urine and fecal<br />

samples with almost equal affinity, allowing for non-invasive testing of this steroid.<br />

typiCAL DAtA<br />

%B/B0<br />

100<br />

90<br />

80<br />

70<br />

60<br />

50<br />

%B/B0<br />

40<br />

30<br />

20<br />

10<br />

0<br />

10 100 1,000 10,000<br />

Estrone Conc. (pg/mL)<br />

reLAteD proDuCts<br />

Estradiol EIA Kits Catalog No. K030-H1/H5 Page 18<br />

Estrone-3- Glucuronide (E1G)EIA Kits Catalog No. K036-H1/H5 Page 20<br />

Progesterone EIA Kits Catalog No. K025-H1/H5 Page 34<br />

Urinary Creatinine Detection Kit Catalog No. K002-H1/H5 Page 14<br />

Hemoglobin Dual Range Detection Kit Catalog No. K013-H1 Page 25<br />

Urea Nitrogen (BUN) Detection Kit Catalog No. K024-H1 Page 41<br />

www.ArborAssays.com | info@ArborAssays.com | Ph: +1.734.677.1774 19<br />

%B/B0<br />

Net OD<br />

0.8<br />

0.7<br />

0.6<br />

0.5<br />

0.4<br />

0.3<br />

0.2<br />

0.1<br />

0.0<br />

Net OD


20<br />

Detectx ®<br />

estrone 3-glucuronide (e1g) <strong>immunoassay</strong> (<strong>eiA</strong>) <strong>kit</strong>s<br />

Catalog No.: K036-H1 (1 Plate) K036-H5 (5 Plate)<br />

FeAtures<br />

f Use Non-Invasive Urine, Fecal Extracts, and TCM<br />

f<br />

f<br />

f<br />

Serum/Plasma Tested in human through fish samples<br />

Fecal Samples Apes, cattle, deer, equids, felids, and ungulates<br />

Samples/Kit 39 in Duplicate in 1 plate <strong>kit</strong>, 231 with 5 plate <strong>kit</strong><br />

iNtroDuCtioN<br />

Estrone-3-glucuronide, C 24 H 30 O 8 , (1,3,5(10)-estratrien-3-ol-17-one glucosiduronate, E1G) is the principle secreted form of<br />

circulating estradiol in mammals. Ovulation is the critical event of each menstrual cycle that occurs during the reproductive<br />

life of healthy females and the ovum can only be fertilized during the short period of time in which it is viable. The three<br />

phases of the menstrual cycle are: (i) an initial phase when there is only a low risk that would enable viable spermatazoa<br />

to survive and reach the ovum, (ii) a phase when the chance of fertilization is at a maximum, the fertile period, and (iii)<br />

a time of absolute infertility when the ovum is no Ionger viable. Clinical studies have indicated the utility of measuring<br />

estrone-3-glucuronide (E1G) and pregnanediol-3a-glucuronide (PDG) in samples of urine or fecal extracts to monitor<br />

ovarian function in females.<br />

estrone-3-glucuronide<br />

The DetectX® Estrone-3-Glucuronide (E1G) Immunoassay <strong>kit</strong> uses a specifically generated antibody to measure E1G and<br />

its metabolites in urine and fecal samples, or in TCM. An E1G standard is provided to generate a standard curve. Standards<br />

or diluted samples are pipetted into a clear microtiter plate coated with an antibody to capture rabbit antibodies. An E1Gperoxidase<br />

conjugate is added to the wells. The binding reaction is initiated by the addition of a polyclonal antibody to<br />

E1G to each well. After a 2 hour incubation the plate is washed and substrate is added. The substrate reacts with the bound<br />

E1G-peroxidase conjugate. After a short incubation, the reaction is stopped and color is read at 450nm.<br />

typiCAL DAtA<br />

%B/B0<br />

100<br />

90<br />

80<br />

70<br />

60<br />

50<br />

%B/B0<br />

40<br />

30<br />

20<br />

10<br />

0<br />

HO<br />

HO<br />

OH<br />

H<br />

OH<br />

reLAteD proDuCts<br />

PGFM Urinary Pregnancy EIA Kits Catalog No. K022-H1/H5 Page 32<br />

Pregnanediol Glucuronide (PDG) EIA Kits Catalog No. K037-H1/H5 Page 32<br />

17ß-Estradiol EIA Kits Catalog No. K030-H1/H5 Page 18<br />

Progesterone EIA Kits Catalog No. K025-H1/H5 Page 34<br />

Estrone EIA Kits Catalog No. K031-H1/H5 Page 19<br />

Urinary Creatinine Detection Kit Catalog No. K002-H1/H5 Page 14<br />

O<br />

%B/B0<br />

Net OD<br />

10 100 1000<br />

Estrone-3-Glucuronide (E1G) Conc. (pg/mL)<br />

1.4<br />

1.2<br />

1.0<br />

0.8<br />

0.6<br />

0.4<br />

0.2<br />

0.0<br />

www.ArborAssays.com | info@ArborAssays.com | Ph: +1.734.677.1774<br />

Net OD


Detectx ®<br />

Formaldehyde Fluorescent Activity <strong>kit</strong><br />

US Patents Catalog No.: K001-F1 (2 Plate)<br />

FeAtures<br />

f Use Measure Formaldehyde in urine, water or TCM<br />

f<br />

f<br />

f<br />

Convenient No extraction, No Chemical Derivatization, 30 minute assay<br />

Species All Species and Samples<br />

Samples/Kit 88 in Duplicate<br />

iNtroDuCtioN<br />

Formaldehyde (methanal), H2C=O, is a colorless, flammable, strong-smelling gas. It is an important industrial chemical<br />

used to manufacture building materials and to produce many household products. In the US approximately 3 x 10 9 Kg<br />

are produced annually. Formaldehyde is commonly used as an industrial fungicide, germicide, and disinfectant, and as<br />

a preservative in mortuaries and medical laboratories. Materials containing formaldehyde can release formaldehyde gas<br />

or vapor into the air. Formaldehyde can also be released by burning wood, kerosene, natural gas, or cigarettes, from<br />

automobile emissions, and from natural processes. Occupational exposure to formaldehyde by inhalation is mainly from<br />

three types of sources: thermal or chemical decomposition of formaldehyde-based resins, formaldehyde emission from<br />

aqueous solutions (for example, embalming fluids), and the production of formaldehyde resulting from the combustion.<br />

Formaldehyde can be toxic, allergenic, and carcinogenic. Because formaldehyde resins are used in many construction<br />

materials it is one of the more common indoor air pollutants.<br />

Formaldehyde<br />

The DetectX® Formaldehyde Detection <strong>kit</strong> is designed to quantitatively measure formaldehyde in a variety of samples<br />

such as urine, water, and buffer solutions. A formaldehyde standard is provided to generate a standard curve for the assay.<br />

The <strong>kit</strong> utilizes a patented non-fluorescent reagent that reacts with formaldehyde in the standards or samples, yielding a<br />

fluorescent product read at 510 nm in a fluorescent plate reader with excitation at 450 nm.<br />

typiCAL DAtA<br />

reLAteD proDuCts<br />

Histone Demethylase Activity Kit Catalog No. K010-F1 Page 26<br />

P450 Demethylating Activity Kit Catalog No. K011-F1 Page 30<br />

Urinary Creatinine Detection Kits Catalog No. K002-H1/H5 Page 14<br />

www.ArborAssays.com | info@ArborAssays.com | Ph: +1.734.677.1774 21


22<br />

Detectx ®<br />

glutathione (gsh) Detection <strong>kit</strong>s<br />

Colorimetric Catalog No.: K006-H1 (4 Plate)<br />

Fluorescent Catalog No.: K006-F1 (1 Plate) K006-F5 (5 Plate)<br />

FeAtures<br />

f Use Measure GSH/GSSG in cells, RBCs, serum, plasma, urine, and tissue<br />

f<br />

f<br />

f<br />

f<br />

Convenient Fluorescent <strong>kit</strong> measures free and total GSH separately in same sample<br />

Sensitive Colorimetric : < 32 pmol/sample Fluorescent: < 2.5 pmol/sample<br />

Samples/Kit Colorimetric: 89 in Duplicate Fluorescent: 39 or 231 in Duplicate<br />

Stability Reagents Stable at 4°C<br />

iNtroDuCtioN<br />

Glutathione (L-γ-glutamyl-L-cysteinylglycine; GSH) is the highest concentration non-protein thiol in mammalian cells and<br />

is present in concentrations of 0.5 – 10 mM. GSH is a tripeptide that contains an unusual peptide linkage between the amine<br />

group of cysteine and the carboxyl group of the glutamate side-chain. It is an antioxidant, preventing damage to important<br />

cellular components caused by reactive oxygen species such as free radicals and peroxides. Glutathione reduces disulfide<br />

bonds formed within cytoplasmic proteins to cysteines by serving as an electron donor. In the process, glutathione is<br />

converted to its oxidized form, glutathione disulfide (GSSG). Glutathione is found mostly in its reduced form, since the<br />

enzyme that reverts it from its oxidized form, glutathione reductase, is constitutive and inducible upon oxidative stress.<br />

The ratio of reduced glutathione to oxidized glutathione within cells is often used as a measure of cellular toxicity.<br />

glutathione<br />

The DetectX® Glutathione Fluorescent <strong>kit</strong> is designed to quantitatively measure glutathione (GSH), followed by oxidized<br />

glutathione (GSSG) using a proprietary non-fluorescent molecule, ThioStar®, that will covalently bind to the free thiol group<br />

on GSH to yield a highly fluorescent product. After adding ThioStar® the fluorescent product formed from GSH is read at 510<br />

nm. Total GSH is measured by the addition of a reaction mixture that converts all the oxidized glutathione, GSSG, into free<br />

GSH, which then reacts with the excess ThioStar®. The Colorimetric <strong>kit</strong> uses a substrate that reacts with the free thiol group<br />

on GSH to yield a highly colored product.<br />

typiCAL DAtA<br />

Fluorescent Colorimetric<br />

reLAteD proDuCts<br />

Glutathione Reductase Activity Kit Catalog No. K009-F1 Page 23<br />

Glutathione S-Tranferase Activity Kit Catalog No. K008-F1 Page 24<br />

AbX Glutathione Antibodies Catalog No. A001/A001F/A001T Page 43<br />

AbX L-Cysteine Antibody Catalog No. A002-50UG Page 43<br />

Glutathione Colorimetric Cuvette Kits Catalog No. K006-H1C-L/H<br />

www.ArborAssays.com | info@ArborAssays.com | Ph: +1.734.677.1774


Detectx ®<br />

glutathione reductase (gr) Fluorescent Activity <strong>kit</strong><br />

FeAtures<br />

f Use Measure GR activity in RBCs, serum, plasma, and cells<br />

f<br />

Convenient Rate or 20 minute end point Fluorescent Kit<br />

f Sensitive 9 µU/mL, World’s Most Sensitive<br />

f<br />

f<br />

Samples/Kit 41 in Duplicate in 20 Minutes<br />

Stability Reagents Stable at 4°C<br />

Catalog No.: K009-F1 (1 Plate)<br />

iNtroDuCtioN<br />

Glutathione reductase (GR) plays an indirect but essential role in the prevention of oxidative damage within the cell by<br />

helping to maintain appropriate levels of intracellular glutathione (GSH). GSH, in conjunction with the enzyme glutathione<br />

peroxidase (GP), is the acting reductant responsible for minimizing harmful hydrogen peroxide cellular levels. The<br />

regeneration of GSH is catalyzed by GR. GR is an ubiquitous 100-120 kDa dimeric flavoprotein that catalyzes the reduction<br />

of oxidized glutathione (GSSG) to reduced glutathione, using ß-nicotinamide dinucleotide phosphate (NADPH) as the<br />

hydrogen donor. NADPH has been suggested to also act as an indirectly operating antioxidant, given its role in the rereduction<br />

of GSSG to GSH and thus maintaining the antioxidative power of glutathione.<br />

glutathione<br />

reductase<br />

reaction<br />

The DetectX® Glutathione Reductase Fluorescent Activity <strong>kit</strong> is designed to quantitatively measure GR activity in a variety<br />

of samples. A GR standard is provided. The <strong>kit</strong> utilizes a proprietary non-fluorescent molecule, ThioStar®, that will covalently<br />

bind to the free thiol group on GSH. Background thiol content is read first after 5 minutes, followed by addition of GSSG<br />

and NADPH which uses the standard or sample GR to convert the oxidized glutathione (GSSG) into free GSH, which then<br />

reacts with the ThioStar® to yield the signal related to GR activity. The signal is read at 510 nm in a fluorescent plate reader<br />

with excitation at 390 nm.<br />

typiCAL DAtA<br />

reLAteD proDuCts<br />

Glutathione Detection Kits Catalog No. K006-F1/F5/H1 Page 22<br />

Glutathione S-Transferase Activity Kit Catalog No. K008-F1 Page 24<br />

AbX Glutathione Antibodies Catalog No. A001/A001F/A001T Page 43<br />

AbX L-Cysteine Antibody Catalog No. A002-50UG Page 43<br />

www.ArborAssays.com | info@ArborAssays.com | Ph: +1.734.677.1774 23


24<br />

Detectx ®<br />

glutathione s-transferase Fluorescent Activity <strong>kit</strong><br />

Catalog No. K008-F1 (1 Plate)<br />

FeAtures<br />

f Use Fluorescent detection of GST Activity<br />

f<br />

f<br />

f<br />

f<br />

Sample Serum, Plasma, and Cell Lysates<br />

Samples/Kit 40 in Duplicate<br />

Convenient 30 Minute End Point or Kinetic Assay<br />

Stability Reagents Stable at 4°C<br />

iNtroDuCtioN<br />

The Glutathione S-Transferase (GST) family of isozymes function to detoxify and neutralize a wide variety of electrophilic<br />

molecules by mediating their conjugation with reduced glutathione. Human GSTs are encoded by 5 gene families, expressing<br />

in almost all tissues as four cytosolic and one microsomal forms. Given its pivotal role in ameliorating oxidative stress/<br />

damage, GST activity has been repeatedly investigated as a biomarker for arthritis, asthma, COPD, and multiple forms<br />

of cancer, as well as an environmental marker. Examination of GST isoforms and activity in human cancers, tumors and<br />

tumor cell lines has revealed the predominance of the acidic pi class. Furthermore, this activity is thought to substantially<br />

contribute to the innate or acquired resistance of specific neoplasms to anticancer therapy.<br />

glutathione<br />

s-transferase<br />

reaction<br />

The DetectX® Glutathione S-Transferase Fluorescent Activity <strong>kit</strong> is designed to quantitatively measure the activity of GST<br />

present in a variety of samples. A GST standard is provided. The <strong>kit</strong> utilizes a non-fluorescent molecule that is a substrate<br />

for the GST enzyme which covalently attaches to glutathione (GSH) to yield a highly fluorescent product. Mixing the<br />

sample or standard with the supplied Detection Reagent and GSH and incubating at room temperature for 30 minutes<br />

yields a fluorescent product which is read at 460 nm in a fluorescent plate reader with excitation at 390 nm.<br />

typiCAL DAtA<br />

reLAteD proDuCts<br />

Glutathione Detection Kits Catalog No. K006-F1/F5/H1 Page 22<br />

Glutathione Reductase Activity Kit Catalog No. K009-F1 Page 23<br />

AbX Glutathione Monoclonal Antibodies Catalog No. A001/A001F/A001T Page 43<br />

AbX L-Cysteine Monoclonal Antibody Catalog No. A002-50UG Page 43<br />

www.ArborAssays.com | info@ArborAssays.com | Ph: +1.734.677.1774


FeAtures<br />

f Sample Blood, RBCs, serum, plasma<br />

f<br />

Rapid 30 Minutes<br />

f Sensitive 20 µg/mL<br />

f<br />

f<br />

Samples/Kit 88 Samples in Duplicate<br />

Stable All Liquid, 4°C Stable Reagents<br />

Detectx ®<br />

hemoglobin Colorimetric Detection <strong>kit</strong><br />

Catalog No: K013-H1 (2 Plate)<br />

iNtroDuCtioN<br />

Hemoglobin (Hgb) is an erythrocyte protein complex comprised of two sets of identical pairs of subunits, each of which<br />

bind an iron-porphyrin group commonly called heme. Heme binds and releases oxygen or carbon dioxide in response to<br />

slight changes in local gas tension. Hemoglobin values are associated with a variety of conditions ranging from anemias<br />

(low Hgb), erythrocytosis (high Hgb), thalassemias (aberrant chain synthesis), and sickling disorders (abnormal complex<br />

shape).<br />

The DetectX® Hemoglobin Detection <strong>kit</strong> is designed to quantitatively measure hemoglobin present in blood and RBCs,<br />

or plasma and serum. A human hemoglobin standard is provided. Standards or diluted samples are pipetted into a clear<br />

microtiter plate and the ready-to-use Hemoglobin Detection Reagent is added. The plate is incubated for 30 minutes at<br />

room temperature and read at 560-580 nm.<br />

typiCAL DAtA<br />

hgb regular range hgb high sensitivity<br />

reLAteD proDuCts<br />

Urinary Creatinine Detection Kits Catalog No. K002-H1/H5 Page 14<br />

www.ArborAssays.com | info@ArborAssays.com | Ph: +1.734.677.1774 25


26<br />

Detectx ®<br />

histone Demethylase (hDm) Fluorescent Activity <strong>kit</strong><br />

Catalog No. K010-F1 (2 Plate) US Patents<br />

FeAtures<br />

f Use Detection of ALL HDM Activity, LSD1 and Jumonji<br />

f<br />

f<br />

f<br />

f<br />

f<br />

Universal Directly Measure Formaldehyde Produced by Demethylation<br />

Sample Cell Lysates<br />

Samples/Kit 89 in Duplicate<br />

Convenient Measure Product in 30 Minutes<br />

Stability All Liquid Reagents Stable at 4°C<br />

iNtroDuCtioN<br />

Formaldehyde is a common by-product formed in the oxidative demethylation of proteins, nucleic acids, and biological<br />

small molecules. Examples of formaldehyde-producing enzymes include DNA demethylases, histone demethylases (HDMs),<br />

and cytochrome P450 enzymes that demethylate drugs and other xenobiotic compounds. HDMs catalyze the site-specific<br />

demethylation of methyl-lysine residues in histones to dynamically regulate chromatin structure, gene expression, and<br />

potentially other genomic functions. Lysine-specific HDMs were first discovered in 2004 and are currently among the most<br />

actively studied formaldehyde-producing enzymes. There are two known classes of HDMs: the flavin adenine dinucleotide<br />

(FAD)-dependent Lysine Specific Demethylase 1 (LSD1) family and the Fe(II)-dependent Jumonji C (JmjC) family. Although<br />

the LSD1 and JmjC HDMs employ different cofactors and catalytic mechanisms, both produce formaldehyde as the product<br />

of the reaction. HDMs have proven difficult to quantitatively assay owing to their relatively low turnover numbers, hindering<br />

our understanding of their kinetic properties, substrate specificities, and reaction mechanisms.<br />

The DetectX® Demethylase Activity <strong>kit</strong> is designed to quantitative the activity of Histone Demethylases. The <strong>kit</strong> is unique in<br />

that the product of these enzymatic demethylation reactions, formaldehyde, is measured directly. No separation or washing<br />

is required. The <strong>kit</strong> has been validated for both LSD1 and JMJD2A Histone Demethylases (HDMs). For in vitro studies, the<br />

HDM reaction should be carried out in our supplied buffers using optimized reaction conditions for the demethylation. For<br />

HDM samples in cells, a special Cell Lysis Buffer is provided. Following the formaldehyde generating reaction the Detection<br />

Reagent is added. After 30 minutes, the fluorescent product is read at 510 nm with excitation at 450 nm.<br />

typiCAL DAtA<br />

LsD1 inhibition profile JmJD2A inhibition profile<br />

reLAteD proDuCts<br />

AbX Histone H3 Antibodies Catalog No. A004/5/6/7 Page 43<br />

AbX LSD1 Antibody Catalog No. A003-100UL Page 43<br />

Tranylcypromine, 10 mM Catalog No. X042-1EA Page 44/45<br />

DNA Hypermethylation Inhibitors Catalog No. P012/P015 Page 44/45<br />

BIX-01294 Catalog No. P018-5MG/25MG Page 44/45<br />

www.ArborAssays.com | info@ArborAssays.com | Ph: +1.734.677.1774


FeAtures<br />

f Sample Urine, Buffer, TCM<br />

f<br />

Rapid 15 Minutes<br />

Detectx ®<br />

hydrogen peroxide (h 2 o 2 ) Fluorescent Detection <strong>kit</strong><br />

f Sensitive < 2 pmole (65 pg) H2O<br />

, World’s Most Sensitive<br />

2<br />

f<br />

Samples/Kit 88 Samples in Duplicate<br />

iNtroDuCtioN<br />

Catalog No.: K034-F1 (2 Plate)<br />

- In biological systems incomplete reduction of O during respiration produces superoxide anion (O ·), which is spontaneously<br />

2 2<br />

- or enzymatically dismutated by superoxide dismutase to H O . Many cells produce low levels of O · and H2O in response to<br />

2 2 2<br />

2<br />

a variety of extracellular stimuli, suchas cytokines (TGF-ß1, TNF-a, and various interleukins), peptide growth factors (PDGF;<br />

EGF, VEGF, bFGF, and insulin), the agonists of heterotrimeric G protein–coupled receptors (GPCR) such as angiotensin II,<br />

thrombin, lysophosphatidic acid, sphingosine 1-phosphate, histamine, and bradykinin, and by shear stress. The addition of<br />

exogenous H O or the intracellular production in response to receptor stimulation affects the function of various proteins,<br />

2 2<br />

including protein kinases, protein phosphatases, transcription factors, phospholipases, ion channels, and G proteins. In<br />

1894, Fenton described the oxidation of tartaric acid by Fe2+ and H O . H O and O may participate in the production of<br />

2 2 2 2 2<br />

singlet oxygen and peroxynitrite and the generation of these species may be concurrent with reactions involving iron, and<br />

under some circumstances they might be important contributors to H O toxicity. A substantial portion of H O lethality<br />

2 2 2 2<br />

involves DNA damage by oxidants generated from iron-mediated Fenton reactions. Damage by Fenton oxidants occurs at<br />

the DNA bases or at the sugar residues. Sugar damage is initiated by hydrogen abstraction from one of the deoxyribose<br />

carbons, and the predominant consequence is eventual strand breakage and base release.<br />

The DetectX® Hydrogen Peroxide Detection Kit is designed to quantitatively measure H 2 O 2 in a variety of samples. A<br />

hydrogen peroxide standard is provided to generate a standard curve for the assay. Samples are mixed with the Fluorescent<br />

Substrate and the reaction initiated by addition of horseradish peroxidase. The reaction is incubated at room temperature<br />

for 15 minutes. The HRP reacts with the substrate in the presence of hydrogen peroxide to convert the colorless substrate<br />

into a fluorescent product. The fluorescent product is read at 590 nm with excitation at 570 nm. Increasing levels of H 2 O 2<br />

cause a linear increase in fluorescent product.<br />

typiCAL DAtA<br />

reLAteD proDuCts<br />

Catalase Fluorescent Activity Kit Catalog No. K033-F1 Page 9<br />

Superoxide Dismutase Activity Kit Catalog No. K028-H1 Page 38<br />

Glutathione Fluorescent Detection Kits Catalog No. K006-F1/F5 Page 22<br />

Glutathione Colorimetric Detection Kit Catalog No. K006-H1 Page 22<br />

Urinary Creatinine Detection Kits Catalog No. K002-H1/H5 Page 14<br />

www.ArborAssays.com | info@ArborAssays.com | Ph: +1.734.677.1774 27


28<br />

Detectx ®<br />

Nitric oxide Colorimetric Detection <strong>kit</strong><br />

Catalog No.: K023-H1 (2 Plate)<br />

FeAtures<br />

f Use Measure Nitrite & Nitrate in Water, Serum, Plasma, Urine, Saliva and TCM<br />

f<br />

f<br />

f<br />

f<br />

f<br />

Accurate Calibrated to NIST Standard Reference material #3185<br />

Sensitive Highest Optical Density of Any Kit<br />

Rapid 5 Minute Nitrite – 30 Minute Total NO<br />

Samples/Kit 88 in Duplicate<br />

Stability Non-Toxic, Stable Reagents at 4°C<br />

iNtroDuCtioN<br />

Nitric oxide (NO) is a diffusible, transient, reactive molecule that has physiological effects in the pM-µM range. Acting<br />

through guanylate cyclase activation, NO is an important regulator of the cardiovascular, nervous, and immunological<br />

systems. NO is bio-available by two routes. It can be endogenously generated by constitutive or induced NOS enzymes<br />

or it can be ingested as nitrates or nitrites for conversion into NO. The reactive nature of nitric oxide allows it to act as a<br />

cytotoxic factor when released during an immune response by macrophages. The reactivity also allows NO to be easily<br />

converted to a toxic radical that can produce nitrosylation damage to cells and DNA. Nitrosylation can be a regulated<br />

post-translational modification in cell signaling. The dynamics of the regulatory/damage facets of NO are major forces in<br />

mitochondrial signaling and dysfunction. NO is linked not only to coronary heart disease, endothelial dysfunctions, erectile<br />

dysfunction, and neurological disorders, but also diabetes, chronic periodontitis, autism and cancer.<br />

The DetectX® Nitric Oxide Detection <strong>kit</strong> is designed to quantitatively measure Nitrate and Nitrite present in a variety<br />

of samples. Nitric Oxide content is derived from the sum of Nitrate ( - NO 3 ) and Nitrite ( - NO 2 ). Both Nitrate and Nitrite<br />

standards are provided. For Nitrite detection, samples are mixed with the Color Reagents A and B and incubated at room<br />

temperature for 5 minutes. The colored product is read at 550–570 nm. Total Nitric Oxide content is measured after the<br />

sample is incubated with Nitrate Reductase and NADH. The reductase, in combination with NADH, reduces Nitrate to<br />

Nitrite. After a 20 minute incubation at room temperature, Color Reagents A and B are added and incubated at room<br />

temperature for 5 minutes. The concentration of Nitrate in the sample is calculated by subtracting the measured Nitrite<br />

concentration from the Total Nitric Oxide concentration for the sample.<br />

typiCAL DAtA<br />

reLAteD proDuCts<br />

Urinary Creatinine Detection Kits Catalog No. K002-H1/H5 Page 14<br />

Urea Nitrogen (BUN) Detection Kit Catalog No. K024-H1 Page 41<br />

Cyclic AMP EIA and CLIA Kits Catalog No. K019-H/C Page 15<br />

Cyclic GMP EIA and CLIA Kits Catalog No. K020-H/C Page 16<br />

Prostaglandin E 2 EIA and CLIA Kits Catalog No. K018-H/HX/C Page 36<br />

www.ArborAssays.com | info@ArborAssays.com | Ph: +1.734.677.1774


FeAtures<br />

f Samples Plasma, urine and milk<br />

f<br />

f<br />

f<br />

f<br />

Fast Three step 2.5 hour assay<br />

Detectx ®<br />

human osteopontin (opN) <strong>immunoassay</strong> (<strong>eiA</strong>) <strong>kit</strong><br />

Catalog No.: K021-H1 (1 Plate)<br />

Trusted By the scientists who published the clinical validation data in Clin. Chem., 2009<br />

Samples/Kit 41 in Duplicate<br />

Stability Reagents Stable at 4°C<br />

iNtroDuCtioN<br />

Osteopontin (OPN) was first described as a secreted, 60 kDa transformation-specific phosphoprotein in 1979. OPN has<br />

been shown to be significant in mineralized tissues, the vascular system, the immune system, kidney and in cancer. OPN<br />

is expressed by a single-copy gene as a 34 kDa nascent protein that is extensively modified by phosphorylation and<br />

glycosylation. OPN is highly conserved in different species and isolated OPNs have a similar number of amino acids, but the<br />

reported size of the secreted protein varies from 44 kDa to 75 kDa, due to differences in post-translational modifications<br />

(PTM). OPN has multiple functions as a key noncollagenous bone matrix protein, a regulator of cytokine production by<br />

macrophages, and in some cancers it has been shown to act as a survival factor. Studies in vitro and in animal models of<br />

cancer have clearly indicated that OPN can function to regulate tumor growth and progression.<br />

The DetectX® Human Osteopontin Immunoassay <strong>kit</strong> is designed to quantitate OPN in plasma, milk and urine samples.<br />

The <strong>kit</strong> uses 2 monoclonal antibodies, one bound to the microtiter plate, the other directly labeled with peroxidase for<br />

detection of bound OPN. The capture monoclonal binds OPN across the single thrombin cleavage site on OPN. The<br />

detection monoclonal binds towards the N-terminal end of OPN. A recombinant human OPN standard, fully modified<br />

with PTMs, is provided. Standards or diluted samples are pipetted into a clear microtiter plate coated with the monoclonal<br />

antibody to capture OPN and the plate is incubated at RT for 1 hour. The plate is washed and peroxidase labeled monoclonal<br />

detection antibody is added. The plate is incubated for another hour at RT. The plate is washed and substrate is added. After<br />

a short incubation, the color reaction is stopped and the intensity read at 450 nm.<br />

typiCAL DAtA<br />

OD<br />

OD<br />

3.5<br />

3<br />

2.5<br />

2<br />

1.5<br />

1<br />

0.5<br />

0<br />

0 5 10 15 20<br />

Osteopontin Conc. (ng/mL)<br />

reLAteD proDuCts<br />

Cyclic AMP Direct EIA and CLIA Kits Catalog No. K019-H/C Page 15<br />

Phosphokinase A (PKA) Activity Kit Catalog No. K027-H1 Page 35<br />

Estrone-3-Glucuronide (E1G) EIA Kits Catalog No. K036-H1/H5 Page 20<br />

Pregnanediol Glucuronide (PDG) EIA Kits Catalog No. K037-H1/H5 Page 33<br />

www.ArborAssays.com | info@ArborAssays.com | Ph: +1.734.677.1774 29


30<br />

Detectx ®<br />

p450 Demethylating Fluorescent Activity <strong>kit</strong><br />

Catalog No.: K011-F1 (2 Plate) US Patents<br />

FeAtures<br />

f Use Measure P450 Demethylating Activity Without Additions to P450 Reaction<br />

f<br />

f<br />

f<br />

f<br />

f<br />

Convenient Run Standard P450 Reaction - Then Measure Formaldehyde Product<br />

Sensitive Fluorescent Assay - Read at 510 nm<br />

Validated 3A4, 2B4, and 2D6 P450s<br />

Samples/Kit 89 Samples in Duplicate<br />

Stability Liquid Reagents Stable at 4°C<br />

iNtroDuCtioN<br />

The cytochromes P450 (P450s) are a superfamily of heme containing enzymes that display tremendous diversity with<br />

regard to substrate specificity and catalytic activity. P450s use a plethora of both compounds as substrates in enzymatic<br />

reactions. Usually they form part of multi-component electron transfer reactions. Catalysis by the eukaryotic P450 enzymes<br />

involves a multistep reaction cycle in which electron transfer is accomplished from a redox partner. The diflavin protein,<br />

NADPH cytochrome P450 reductase (reductase), contains both FAD and FMN and can transfer both electrons needed for<br />

the catalytic cycle. In some P450 reactions, the second electron of the reaction cycle also can be delivered by cytochrome<br />

b5. The P450 enzymes and cofactors of the mammalian drug-metabolizing system are embedded in the membrane of the<br />

endoplasmic reticulum. The P450s play a crucial role in the development of new drug entities as drug-drug interactions<br />

commonly arise from the inhibition of cytochrome P450 activities. Lipid plays an important role in the reconstitution of<br />

P450-dependent activities after protein purification. Most in vitro studies for the reconstitution of P450 activities use<br />

DLPC as the lipid component. The reconstitution of enzymatic activity involves a concentrated incubation of P450, its<br />

redox partners (NADPH and reductase), and lipid. The reported preincubation conditions vary significantly.<br />

The DetectX® P450 Activity <strong>kit</strong> is designed to quantitatively measure the enzymatic activity of formaldehyde-producing<br />

enzymes such as Cytochrome P450s. The <strong>kit</strong> is unique in that the fluorescent substrate (FDR) is not involved in the multicomponent<br />

P450 reaction, but measures the product of the demethylation, formaldehyde. No separation or washing<br />

is required. The <strong>kit</strong> has been validated for several P450 systems and should work with any biological system that is<br />

producing formaldehyde as a product of demethylation. Following the P450 NADPH-induced reaction, the generation of<br />

formaldehyde can be stopped by addition of a suitable inhibitor, or the supplied stop solution of acetic acid. The FDR is<br />

added to all the wells. After a short incubation at 37°C for 30 minutes, the fluorescent product is read at 510 nm.<br />

typiCAL DAtA<br />

www.ArborAssays.com | info@ArborAssays.com | Ph: +1.734.677.1774


FeAtures<br />

f Use Measure Pd Contamination in APIs in 30 Minutes<br />

f<br />

f<br />

f<br />

f<br />

f<br />

Rapid Estimate Pd Scavenger Methods with HTS<br />

pdx <br />

palladium Api Fluorescent screening <strong>kit</strong><br />

Patent Pending Catalog No. K007-F1 (1 Plate)<br />

Adaptable APIs in HCl, NMP, DMSO, DMF, MeCN, EtOH or Toluene<br />

Sensitive Low ppm Pd Detectable<br />

Samples/Kit 41 in Duplicate<br />

Stability All Liquid Reagents Stable at RT<br />

iNtroDuCtioN<br />

Many new synthetic transformations have been developed that use palladium (Pd) compounds for the catalysis of carboncarbon<br />

and carbon-heteroatom coupling reactions. These reactions have found increased popularity for pharmaceutical<br />

processes as they utilize a wide-range of functional groups and can therefore be used to build complicated molecules.<br />

However, palladium-catalyzed reactions present a problem in that the palladium can often be retained in the isolated<br />

product. The LD50 values are very dependent on the physical form of the Pd compounds or catalysts used. For rats, mice,<br />

or rabbits, water soluble PdCl 2 administered intravenously gave a LD50 of 3 mg/kg body weight while relatively insoluble<br />

PdO given orally gave an LD50 >4900 mg/kg body weight. Current European Agency for the Evaluation of Medicinal<br />

Products regulations limit all platinum group (Pt, Pd, Ir, Rh, Ru, Os) metal contamination (as a group) to less than 5 ppm. A<br />

variety of methods are available for removing Pd from active pharmaceutical ingredients (APIs). These include adsorption<br />

with trimercaptotriazine N-acetylcysteine, such as the Isolute resins (Biotage). Other adsorptive methods include activated<br />

carbon, ion exchange type resins, fiberous materials such as Smopex (Johnson Matthey), and silica bound active resins<br />

from SiliCycle. Typical chemical purification methods such as crystallization can also be used to lower the concentration<br />

of Pd in the final product, especially when combined with additives such as the sulfur containing ligands, N-acetylcysteine<br />

and thiourea, and with phosphines to keep the Pd in the mother liquor. The standard methods of quantifying palladium in<br />

APIs are atomic absorption analysis, x-ray fluorescence, and plasma emission spectroscopy. ICP-MS is typically used both<br />

during synthesis and for final QC on drug molecules.<br />

PdX® Palladium (Pd) Detection <strong>kit</strong> is designed to allow the rapid determination of the relative amounts of Pd present<br />

in active pharmaceutical ingredient (API) scavenging steps. The <strong>kit</strong> uses a patent-pending, non-fluorescent detection<br />

molecule that, under reducing conditions, palladium cleaves to yield a brightly fluorescent product which is read at 520<br />

nm with excitation at 485 nm.<br />

typiCAL DAtA<br />

www.ArborAssays.com | info@ArborAssays.com | Ph: +1.734.677.1774 31


32<br />

Detectx ®<br />

pgFm (13,14-Dihydro-15-keto-prostaglandin F 2a ) <strong>eiA</strong> <strong>kit</strong>s<br />

Catalog No. K022-H1 (1 Plate) K022-H5 (5 Plate) Patent Pending<br />

FeAtures<br />

f Use Non-Invasive Urine or Fecal Extracts, Plus Serum, Plasma and TCM<br />

f<br />

f<br />

f<br />

f<br />

Serum/Plasma Tested in human through fish samples<br />

Urine/Fecal Apes, cattle, deer, equids, felids, and ungulates<br />

Samples/Kit 39 in Duplicate in 1 plate <strong>kit</strong>, 231 with 5 plate <strong>kit</strong><br />

Stability All Liquid Reagents Stable at 4°C<br />

iNtroDuCtioN<br />

In many species, uterine and placental Prostaglandin F 2a (PGF 2a ) is involved in the regulation of reproductive and pregnancyrelated<br />

processes such as embryonic development, initiation of parturition, and resumption of ovarian activity. In domestic<br />

ruminants, uterine tissue is a primary source of PGF 2a , and secretion of uterine PGF 2a is a key regulator for the cyclical<br />

regression of the corpus luteum. Prostaglandin F 2a is metabolized to PGFM (13,14-dihydro-15-keto-PGF 2a ) during the first<br />

passage through the lungs. PGFM has a longer half-life in peripheral circulation than PGF 2a and has been applied as a<br />

useful analytical marker of PGF 2a . PGFM is a useful non-invasive marker of pregnancy when measured in both urine and<br />

fecal samples (see below for Iberian Lynx data). It has been shown to be a precise, practical method for this application<br />

in these matrices. Parallel courses were obtained when comparing urinary and fecal PGFM in a variety of felids and<br />

other species, and only a simple dilution of fecal extracts is necessary prior to analysis. Fecal PGFM analyses may allow<br />

pregnancy diagnosis in captive and free-ranging felids. Recent evidence has suggested that PGFM may also be a useful<br />

pregnancy marker in some other non-felid species.<br />

%B/B0<br />

100<br />

90<br />

80<br />

70<br />

60<br />

50<br />

%B/B0<br />

40<br />

30<br />

20<br />

10<br />

0<br />

13,14-dihydro-15-keto-pgF 2a (pgFm)<br />

The DetectX® 13,14-dihydro-15-keto-PGF 2a (PGFM) Immunoassay <strong>kit</strong> is a patent pending assay from Dr. Martin Dehnhard at<br />

Leibniz Institute for Zoo & Wildlife Research in Berlin, licensed exclusively and designed to quantitatively measure PGFM<br />

present in fecal extracts, urine, serum and plasma samples. A PGFM standard is provided. Standards or diluted samples<br />

are pipetted into a clear microtiter plate coated with an antibody to capture rabbit IgG. A PGFM-peroxidase conjugate is<br />

added. The binding reaction is initiated by the addition of a rabbit polyclonal antibody highly specific to PGFM to each<br />

well. After an hour incubation, the plate is washed and substrate is added. The substrate reacts with the bound PGFMperoxidase<br />

conjugate. After a short incubation, the reaction is stopped and the intensity is read at 450 nm.<br />

typiCAL DAtA<br />

%B/B0<br />

Net OD<br />

10 100 1,000<br />

0<br />

10,000<br />

PGFM Conc. (pg/mL)<br />

0.50<br />

0.45<br />

0.40<br />

0.35<br />

0.30<br />

0.25<br />

0.20<br />

0.15<br />

0.10<br />

0.05<br />

Net OD<br />

Net OD<br />

pregnant and pseudo-pregnant iberian Lynx data<br />

reLAteD proDuCts<br />

Progesterone EIA Kits Catalog No. K025-H1/H5 Page 34<br />

Estradiol EIA Kits Catalog No. K030-H1/H5 Page 18<br />

Estrone 3-Glucuronide EIA Kits Catalog No. K031-H1/H5 Page 20<br />

Pregnanediol Glucuronide (PDG) EIA Kits Catalog No. K037-H1/H5 Page 33<br />

Ceruloplasmin Colorimetric Activity Kit Catalog No. K035-H1 Page 10<br />

Urinary Creatinine Detection Kit Catalog No. K002-H1/H5 Page 14<br />

www.ArborAssays.com | info@ArborAssays.com | Ph: +1.734.677.1774


Detectx ®<br />

pregnanediol glucuronide (pDg) <strong>immunoassay</strong> (<strong>eiA</strong>) <strong>kit</strong>s<br />

FeAtures<br />

f Use Measure 11- and 3-Progesterone Derivatives<br />

f<br />

f<br />

f<br />

Samples Urine, Fecal Extracts and TCM<br />

Samples/Kit 41 in Duplicate in 1 plate <strong>kit</strong>, 233 with 5 plate <strong>kit</strong><br />

Stability All Liquid Reagents Stable at 4°C<br />

Catalog No.: K037-H1 (1 Plate) K037-H5 (5 Plate)<br />

iNtroDuCtioN<br />

Pregnanediol Glucuronide, C 27 H 44 O 8 , also known as PDG (5ß-Pregnnan-3a,20a-diol 3-glucosiduronate) is the major<br />

metabolite of progesterone. Progesterone is the hormone involved in the female menstrual cycle, gestation and<br />

embryogenesis of humans and other species. Progesterone belongs to a class of hormones called progestogens, and is<br />

the major naturally occurring human progestogen. Progesterone is an essential regulator of human female reproductive<br />

function in the uterus, ovary, mammary gland and brain, and plays an important role in non-reproductive tissues such<br />

as the cardiovascular system, bone and the central nervous system. Progesterone action is conveyed by two isoforms<br />

of the nuclear progesterone receptor (PR), PRA and PRB. PRA and B are expressed in a variety of normal breast tissue<br />

from humans, rats and mice and is also expressed in breast cancer cells. Progesterone also has neurotrophic roles in the<br />

peripheral nervous system as it activates the growth and maturation of axons and stimulates the repair and replacement<br />

of myelin sheaths in regenerating nerve fibres.<br />

pregnanediol glucuronide<br />

The DetectX® PDG Immunoassay <strong>kit</strong> measures PDG present in urine and extracted dried fecal samples. A PDG standard is<br />

provided for the assay. Standards or diluted samples are pipetted into a coated clear microtiter plate. A PDG-peroxidase<br />

conjugate is added to the wells. The binding reaction is initiated by the addition of a polyclonal antibody to PDG. After<br />

a 2 hour incubation the plate is washed and substrate is added. The substrate reacts with the bound PDG-peroxidase<br />

conjugate. After a short incubation the intensity of the generated color is measured at 450 nm.<br />

typiCAL DAtA<br />

%B/B0<br />

100<br />

90<br />

80<br />

70<br />

60<br />

50<br />

%B/B0<br />

40<br />

30<br />

20<br />

10<br />

0<br />

HO<br />

100 1,000 10,000 100,000<br />

Pregnanediol Glucuronide (PDG) Conc. (pg/mL)<br />

reLAteD proDuCts<br />

Progesterone EIA Kits Catalog No. K025-H1/H5 Page 34<br />

Estradiol EIA Kits Catalog No. K030-H1/H5 Page 18<br />

Estrone EIA Kits Catalog No. K031-H1/H5 Page 19<br />

Estrone 3-Glucuronide EIA Kits Catalog No. K031-H1/H5 Page 20<br />

PGFM EIA Kits Catalog No. K022-H1/H5 Page 32<br />

Ceruloplasmin Colorimetric Activity Kit Catalog No. K035-H1 Page 10<br />

Urinary Creatinine Detection Kit Catalog No. K002-H1/H5 Page 14<br />

HO<br />

%B/B0<br />

Net OD<br />

OH<br />

H<br />

OH<br />

O<br />

www.ArborAssays.com | info@ArborAssays.com | Ph: +1.734.677.1774 33<br />

2.5<br />

2<br />

1.5<br />

1<br />

0.5<br />

0<br />

Net OD


34<br />

Detectx ®<br />

progesterone <strong>immunoassay</strong> (<strong>eiA</strong>) <strong>kit</strong>s<br />

Catalog No.: K025-H1 (1 Plate) K025-H5 (5 Plate)<br />

FeAtures<br />

f Use Measure 11- and 3-Hydroxy Progesterones<br />

f<br />

f<br />

f<br />

Samples Urine and Fecal Extracts and TCM<br />

Samples/Kit 39 in Duplicate in 1 plate <strong>kit</strong>, 231 with 5 plate <strong>kit</strong><br />

Stability All Liquid Reagents Stable at 4°C<br />

iNtroDuCtioN<br />

Progesterone, C 21 H 30 O 2 , also known as P4 (pregn-4-ene-3,20-dione) is a C-21 steroid hormone involved in the female<br />

menstrual cycle, gestation and embryogenesis of humans and other species. Progesterone belongs to a class of hormones<br />

called progestogens, and is the major naturally occurring human progestogen. Progesterone is an essential regulator of<br />

human female reproductive function in the uterus, ovary, mammary gland and brain, and plays an important role in nonreproductive<br />

tissues such as the cardiovascular system, bone and the central nervous system. Progesterone action is<br />

conveyed by two isoforms of the nuclear progesterone receptor (PR), PRA and PRB. PRA and B are expressed in a variety<br />

of normal breast tissue from humans, rats and mice and is also expressed in breast cancer cells. Progesterone also has<br />

neurotrophic roles in the peripheral nervous system as it activates the growth and maturation of axons and stimulates the<br />

repair and replacement of myelin sheaths in regenerating nerve fibres.<br />

progesterone<br />

The DetectX® Progesterone Immunoassay <strong>kit</strong> measures Progesterone present in urine and extracted dried fecal samples. A<br />

progesterone standard is provided for the assay. Standards or samples are pipetted into a coated clear microtiter plate. A<br />

progesterone-peroxidase conjugate is added to the wells. The binding reaction is initiated by the addition of a monoclonal<br />

antibody to progesterone. After a 2 hour incubation the plate is washed and substrate is added. The substrate reacts with<br />

the bound progesterone-peroxidase conjugate. After a short incubation the intensity of the generated color is measured<br />

at 450 nm.<br />

typiCAL DAtA<br />

reLAteD proDuCts<br />

Pregnanediol Glucuronide (PDG) EIA Kits Catalog No. K037-H1/H5 Page 33<br />

Estrone-3-Glucuronide (E1G) EIA Kits Catalog No. K036-H1/H5 Page 20<br />

Testosterone EIA Kits Catalog No. K032-H1/H5 Page 39<br />

Estradiol EIA Kits Catalog No. K030-H1/H5 Page 18<br />

Estrone EIA Kits Catalog No. K031-H1/H5 Page 19<br />

Urinary Creatinine Detection Kit Catalog No. K002-H1/H5 Page 14<br />

www.ArborAssays.com | info@ArborAssays.com | Ph: +1.734.677.1774


Detectx ®<br />

phosphokinase A (pkA) Colorimetric Activity <strong>kit</strong><br />

FeAtures<br />

f Use Measure PKA Activity in a Variety of Samples<br />

f<br />

f<br />

f<br />

f<br />

Sensitive Most Sensitive Kit<br />

Rapid 3 Hour Assay<br />

Convenient Simple 3 Step Protocol<br />

Samples/Kit 42 Samples in Duplicate<br />

Catalog No.: K027-H1 (1 Plate)<br />

iNtroDuCtioN<br />

The expressed PKA holoenzyme, compromising of two catalytic (C) and two regulatory (R) subunits, is activated when<br />

cAMP levels rise following stimulation of Gs protein-coupled receptors and adenylyl cyclase. The phosphorylation of<br />

specific substrates by the C subunit of activated PKA is regulated by the subcellular localization of the enzyme through<br />

the binding to the scaffolding A kinase-anchoring proteins (AKAPs). In its inactive state, the pseudosubstrate sequences<br />

on the R subunits stop the activity of the C subunits. Upon binding of cAMP to the R subunits, the active monomeric C<br />

subunits are released. PKA shares substrate specificity with Akt (PKB) and PKC. Substrates that are phosphorylated by<br />

PKA are Bad (Ser 155 ), CREB (Ser 133 ), and GSK-3 (GSK-3a Ser 21 and GSK -3ß Ser 9 ). PKA is a pivotal kinase involved in cancer,<br />

vasodilation, metabolic processes, etc.<br />

The DetectX® PKA Activity <strong>kit</strong> is designed to quantitatively measure the enzymatic activity of PKA in samples. The <strong>kit</strong><br />

utilizes a plate bound PKA substrate that is phosphorylated in the presence of PKA and ATP. Following an incubation, the<br />

plate is washed and an antibody specific for the phosphorylated substrate and the peroxidase-labeled detection conjugate<br />

are added. The excess antibody and conjugate are washed out and substrate added. The substrate reacts with the bound<br />

peroxidase conjugate. After a short incubation the intensity of the generated color is measured at 450 nm.<br />

typiCAL DAtA<br />

Net OD (450nm)<br />

1.4<br />

1.2<br />

1.0<br />

0.8<br />

0.6<br />

OD (450 nm)<br />

0.4<br />

0.2<br />

0.0<br />

0 5 10 15 20 25<br />

PKA Activity (U/mL)<br />

reLAteD proDuCts<br />

Cyclic AMP EIA and CLIA Kits Catalog No. K019-H/C Page 15<br />

Cyclic GMP EIA and CLIA Kits Catalog No. K020-H/C Page 16<br />

Prostaglandin E 2 EIA and CLIA Kits Catalog No. K018-H/HX/C Page 36<br />

IBMX, Pan-specific PDE Inhibitor Catalog No. P019-100MG/1G Page 44/45<br />

www.ArborAssays.com | info@ArborAssays.com | Ph: +1.734.677.1774 35


36<br />

Detectx ®<br />

prostaglandin e 2 (pge 2 ) <strong>immunoassay</strong> (<strong>eiA</strong> and CLiA) <strong>kit</strong>s<br />

Regular Format Catalog No: K018-H1 (1 Plate) K018-H5 (5 Plate)<br />

High Sensitivity Catalog No: K018-HX1 (1 Plate) K018-HX5 (5 Plate)<br />

Chemiluminescent Catalog No: K018-C1 (1 Plate) K018-C5 (5 Plate)<br />

FeAtures<br />

f Use Measure PGE in cells, saliva, urine, serum, plasma and tissue<br />

2<br />

f 3 Formats Regular: 1,000-31.25 High Sensitivity: 400-12.5 CLIA: 320-5 pg/mL<br />

f<br />

f<br />

Sensitive Regular : < 3 pg High Sensitivity: < 1.1 pg CLIA:


Detectx ®<br />

retinol binding protein (rbp) <strong>immunoassay</strong> (<strong>eiA</strong>) <strong>kit</strong>s<br />

FeAtures serum <strong>kit</strong> urinary <strong>kit</strong><br />

f Use Vitamin A Deficiency Kidney Function<br />

f<br />

f<br />

f<br />

f<br />

Range (ng/mL) 78.1-5,000 3.9-1,000<br />

Sample Serum or Plasma Urine<br />

Serum Catalog No.: K004-H1 (1 Plate)<br />

Urinary Catalog No.: KU04-H1 (1 Plate)<br />

Species Human, Rat, Dog, Monkey Human, Rat, Dog, Monkey<br />

Time to Answer 75 Minutes 90 Minutes<br />

iNtroDuCtioN<br />

Retinol binding protein (RBP) is from a family of structurally related proteins that bind small hydrophobic molecules such as<br />

bile pigments, steroids, odorants, etc. RBP is a 21 kDa highly conserved, single-chain glycoprotein, consisting of 182 amino<br />

acids with 3 disulfide bonds, that has a hydrophobic pocket which binds retinol (vitamin A).<br />

The RBP Serum EIA <strong>kit</strong> is used to assess Vitamin A Deficiency (VAD) as RBP levels in the peripheral circulation decrease in<br />

response to decreased amounts of vitamin A. The RBP Urinary EIA <strong>kit</strong> is specially formulated to measure urine RBP as it has<br />

also been shown to be a useful marker for renal function. RBP is totally filtered by the glomeruli and reabsorbed by proximal<br />

tubules. Urinary RBP is used to study renal function in heart or kidney transplant recipients, type 1 and 2 diabetics, and in<br />

people exposed to uranium from mining operations.<br />

The DetectX® Immunoassay <strong>kit</strong>s use a RBP standard to generate a standard curve. Standards or diluted samples are pipetted<br />

into a clear coated microtiter plate. A RBP-peroxidase conjugate is added to the standards and samples in the wells. The<br />

binding reaction is initiated by the addition of the RBP polyclonal antibody to each well. After the primary incubation the<br />

plate is washed and substrate is added. The substrate reacts with the bound RBP-peroxidase conjugate. The color reaction<br />

is stopped and the intensity is read at 450 nm.<br />

typiCAL DAtA<br />

reLAteD proDuCts<br />

Serum Creatinine Detection Kits Catalog No. KB02-H1/H2/H1D Page 14<br />

Cystatin C EIA Kit Catalog No. K012-H1 Page 17<br />

Glutathione Detection Kits Catalog No. K006-F1/F5/H1 Page 22<br />

Nitric Oxide Detection Kit Catalog No. K023-H1 Page 28<br />

www.ArborAssays.com | info@ArborAssays.com | Ph: +1.734.677.1774 37


38<br />

Detectx ®<br />

superoxide Dismutase (soD) Colorimetric Activity <strong>kit</strong><br />

Catalog No.: K028-H1 (2 Plate)<br />

FeAtures<br />

f Use Measure SOD Activity<br />

f<br />

f<br />

f<br />

Sample Serum, Urine, and buffer samples<br />

Species Human and other mammalian species<br />

Samples/Kit 89 in Duplicate<br />

iNtroDuCtioN<br />

·- · Short-lived and highly reactive oxygen species (ROS) such as O (superoxide), OH (hydroxyl radical), and H2O (hydrogen<br />

2<br />

2<br />

peroxide) are continuously generated in vivo. The cellular levels of ROS are controlled by antioxidant enzymes and small<br />

molecule antioxidants. The major antioxidant enzymes, superoxide dismutases (SODs), including copper-zinc superoxide<br />

dismutase (Cu/ZnSOD), manganese superoxide dismutase (MnSOD) and extracellular superoxide dismutase (EC-SOD), all<br />

play a critical roles in scavenging O2· . Decreased SOD activity results in elevated level of superoxide which in turn leads<br />

to decreased NO and increased peroxynitrite concentrations. The major intracellular SOD is a 32-kD copper and zinc<br />

containing homodimer (Cu/Zn SOD). The mitochondrial SOD (MnSOD) is a manganese-containing 93-kD homotetramer<br />

that is synthesized in the cytoplasm and translocated to the inner matrix of mitochondria. EC-SOD is the primary extracellular<br />

SOD enzyme and is highly expressed in many organs. Increased SOD activity levels are seen in Downs Syndrome, while<br />

decreased activity is seen in diabetes, Alzheimer’s disease, rheumatoid arthritis, Parkinson’s disease, uremic anemia,<br />

atherosclerosis, some cancers, and thyroid dysfunction.<br />

The DetectX® Superoxide Dismutase (SOD) Activity <strong>kit</strong> is designed to quantitatively measure SOD activity in a variety<br />

of samples. A bovine erythrocyte SOD standard is provided. Samples are mixed with a Detection Reagent and Reaction<br />

Mixture, then incubated at room temperature for 20 minutes. Xanthine and xanthine oxidase in the Reaction Mixture generate<br />

superoxide in the presence of oxygen, which converts a colorless substrate in the Detection Reagent into a yellow colored<br />

product. The colored product is read at 450 nm. Increasing levels of SOD in the samples causes a decrease in superoxide<br />

concentration and a reduction in yellow product.<br />

typiCAL DAtA<br />

OD<br />

OD<br />

0.7<br />

0.6<br />

0.5<br />

0.4<br />

0.3<br />

0.2<br />

0.1<br />

0<br />

0.1 1 10 100<br />

SOD Activity (U/mL)<br />

reLAteD proDuCts<br />

Urinary Creatinine Detection Kits Catalog No. K002-H1/H5 Page 14<br />

Hydrogen Peroxide Detection Kit Catalog No. K034-F1 Page 27<br />

Hemoglobin Dual Range Detection Kit Catalog No. K013-H1 Page 25<br />

Urea Nitrogen (BUN) Detection Kit Catalog No. K024-H1 Page 41<br />

www.ArborAssays.com | info@ArborAssays.com | Ph: +1.734.677.1774


FeAtures<br />

f Use Measure Testosterone and Dihydrotestosterone<br />

f<br />

f<br />

f<br />

Samples Urine, Fecal Extracts and TCM<br />

Samples/Kit 39 in Duplicate in 1 plate <strong>kit</strong>, 231 with 5 plate <strong>kit</strong><br />

Stability All Liquid Reagents Stable at 4°C<br />

Detectx ®<br />

testosterone <strong>immunoassay</strong> (<strong>eiA</strong>) <strong>kit</strong>s<br />

Catalog No.: K032-H1 (1 Plate) K032-H5 (5 Plate)<br />

iNtroDuCtioN<br />

Testosterone, C 19 H 28 O 2 , (4-Androsten-17ß-ol-3-one) is a steroid hormone from the androgen group and is found in mammals,<br />

reptiles, birds, and other vertebrates. In mammals, testosterone is primarily secreted in the testes of males and the ovaries<br />

of females, although small amounts are also secreted by the adrenal glands. It is the principal male sex hormone and an<br />

anabolic steroid. In men, testosterone plays a key role in the development of male reproductive tissues such as the testis<br />

and prostate, and promoting secondary sexual characteristics such as increased muscle, bone mass, and body hair. In the<br />

absence of testosterone stimulation, spermatogenesis does not proceed beyond the meiosis stage. In addition, testosterone<br />

is essential for health and well-being as well as the prevention of osteoporosis. Testosterone plays a significant role in glucose<br />

homeostasis and lipid metabolism. The metabolic syndrome is a clustering of risk factors predisposing to type 2 diabetes<br />

mellitus, atherosclerosis and cardiovascular morbidity and mortality. Cross-sectional epidemiological studies have reported<br />

a direct correlation between plasma testosterone and insulin sensitivity, and low testosterone levels are associated with an<br />

increased risk of type 2 diabetes, dramatically illustrated by androgen deprivation in men with prostate carcinoma.<br />

testosterone<br />

The DetectX® Testosterone Immunoassay <strong>kit</strong> measures Testosterone present in urine and extracted dried fecal samples. A<br />

testosterone standard is provided for the assay. Standards or samples are pipetted into a coated clear microtiter plate. A<br />

testosterone-peroxidase conjugate is added to the wells. The binding reaction is initiated by the addition of a polyclonal<br />

antibody to testosterone. After a 2 hour incubation the plate is washed and substrate is added. The substrate reacts with the<br />

bound testosterone-peroxidase conjugate. After a short incubation the intensity of the generated color is measured at 450 nm.<br />

typiCAL DAtA<br />

reLAteD proDuCts<br />

Osteopontin EIA Kit Catalog No. K021-H1 Page 29<br />

PGFM Urinary Pregnancy EIA Kits Catalog No. K022-H1/H5 Page 32<br />

Pregnanediol Glucuronide (PDG) EIA Kits Catalog No. K037-H1/H5 Page 33<br />

17ß-Estradiol EIA Kits Catalog No. K030-H1/H5 Page 18<br />

Progesterone EIA Kits Catalog No. K025-H1/H5 Page 34<br />

Estrone EIA Kits Catalog No. K031-H1/H5 Page 19<br />

Urinary Creatinine Detection Kit Catalog No. K002-H1/H5 Page 14<br />

Hemoglobin Dual Range Detection Kit Catalog No. K013-H1 Page 19<br />

Urea Nitrogen (BUN) Detection Kit Catalog No. K024-H1 Page 19<br />

www.ArborAssays.com | info@ArborAssays.com | Ph: +1.734.677.1774 39


40<br />

Detectx ®<br />

thiol Fluorescent Detection <strong>kit</strong><br />

Catalog No.: K005-F1 (1 Plate)<br />

FeAtures<br />

f Use Measure Thiol content of Recombinant Proteins<br />

f<br />

f<br />

f<br />

f<br />

f<br />

Adaptable Measure Protein SH in 6M GuHCl Buffers<br />

Sensitive < 0.5 pmol Thiol/well<br />

Rapid 30 Minute Assay<br />

Samples/Kit 39 in Duplicate<br />

Stability Non-Toxic, Reagents Stable at 4°C<br />

iNtroDuCtioN<br />

Free thiols in biological systems have important roles. Oxidatively modified thiol groups of cysteine residues are known<br />

to modulate the activity of a growing number of proteins. One of the most pressing problems with this approach is to<br />

accurately determine the extent of modification of specific amino acids, such as cysteine residues, in a complex protein<br />

sample, especially in the presence of chaotropic agents such as guanidine hydrochloride.<br />

The DetectX® Thiol <strong>kit</strong> is designed to quantitatively measure thiol groups generated or present in biological samples.<br />

A N-Acetyl Cysteine standard is provided. Samples and standards are pipetted into a black microtiter plate. Mixing the<br />

samples or standards with ThioStar® and incubating at room temperature, the fluorescent product is read at 510 nm with<br />

excitation at 390 nm.. This assay is adaptable for measurement in higher density plate formats.<br />

typiCAL DAtA<br />

Comparison to ellman’s effect of guanidine hCl<br />

reLAteD proDuCts<br />

ThioStar® Detection Reagent Catalog No. L002 Page 42<br />

www.ArborAssays.com | info@ArborAssays.com | Ph: +1.734.677.1774


FeAtures<br />

f Sample Serum, plasma, urine, saliva<br />

f<br />

Rapid 30 Minutes<br />

f Sensitive 30 µg/dL<br />

f<br />

Sample/Kit 88 Samples in Duplicate<br />

Detectx ®<br />

urea Nitrogen (buN) Detection <strong>kit</strong><br />

Catalog No.: K024-H1 (2 Plate)<br />

iNtroDuCtioN<br />

Urea is a by-product of protein metabolism by the liver, and is therefore removed from the blood by the kidneys. Urea<br />

freely filters through the glomerulous, but is reabsorbed by the renal tubules in a flow-dependent fashion. The higher the<br />

flow rate, the greater amount of urea nitrogen is cleared from circulation and eliminated through the kidneys. As a result,<br />

the level of circulating urea nitrogen, along with serum creatinine, serves as a primary measure of kidney function. Normal<br />

adult Blood Urea Nitrogen (BUN) levels should be between 7 and 21 mg urea nitrogen per 100 mL blood (mg/dL).<br />

Azotemia, poor kidney function, will cause elevated BUN levels (≥ 50 mg/dL) and is associated with acute kidney failure<br />

or injury, severe acute pancreatitis, congestive heart failure or gastrointestinal bleeding. Azotemia also can occur with<br />

dehydration, as a result of alcohol abuse, or high protein diets. Lower than expected BUN levels are usually not clinically<br />

predictive, but are primarily associated with liver disease or malnutrition, including malabsorption and low protein diets.<br />

Urine and saliva are considered to be acceptable non-invasive samples for measurement of urea nitrogen.<br />

The DetectX® Urea Nitrogen (also called BUN) Detection Kit is designed to quantitatively measure urea nitrogen in a variety<br />

of samples. A urea nitrogen standard calibrated to NIST reference materials is provided to generate a standard curve<br />

for the assay. Samples are mixed with Color Reagents A and B and incubated at room temperature for 30 minutes. The colored<br />

product is read at 450 nm. The concentration of urea nitrogen in the sample is calculated. The results are expressed<br />

in terms of mg/dL urea nitrogen. If samples are to be expressed in terms of mg/dL urea, the data can be converted using<br />

the multiplier 2.14.<br />

typiCAL DAtA<br />

reLAteD proDuCts<br />

Urinary Creatinine Detection Kits Catalog No. K002-H1/H5 Page 14<br />

Serum Creatinine Detection Kits Catalog No. KB02-H1/H2/H1D Page 14<br />

Cystatin C EIA Kit Catalog No. K012-H1 Page 17<br />

Urinary RBP EIA Kit Catalog No. KU04-H1 Page 37<br />

Hemoglobin Colorimetric Detection Kit Catalog No. K013-H1 Page 25<br />

www.ArborAssays.com | info@ArborAssays.com | Ph: +1.734.677.1774 41


42<br />

thiostar ® thiol Detection reagent<br />

Catalog No.: L002-50UG/100UG/250UG/500UG<br />

FeAtures<br />

f Rapid Measure low concentration of thiols<br />

f<br />

Fluorescent Ideal for high throughput applications<br />

iNtroDuCtioN<br />

The ThioStar® Thiol Detection Reagent allows users to accurately determine the extent of free thiol content in samples.<br />

ThioStar is converted to a brightly fluorescent product upon reaction with thiols in the sample. The thiol in the sample can<br />

either be one that is generated by a reaction, such as the end product of an enzymatic reaction, or can be the cysteine<br />

content of the protein to be measured. ThioStar is used by simply diluting the ThioStar reagent in dry DMSO and adding it to<br />

the thiol sample in buffer. ThioStar will work in almost any buffer. ThioStar should be stored desiccated.<br />

thiostar® kinetics excitation spectra emission spectra<br />

recombinant obelin<br />

Catalog No.: L001-50UG/100UG<br />

FeAtures<br />

f Rapid Measure low concentrations of calcium<br />

f<br />

f<br />

60,000<br />

50,000<br />

40,000<br />

30,000<br />

FLu<br />

20,000<br />

10,000<br />

0<br />

0 10 20 30 40 50 60<br />

time (mins)<br />

Bioluminescent Ideal for high throughput applications<br />

120,000<br />

100,000<br />

60,000<br />

FLu<br />

Sensitive No background signal<br />

iNtroDuCtioN<br />

Recombinant bioluminescent photoprotein from Obelia<br />

longissima. The photoprotein is approximately 21,000<br />

molecular weight and contains the luminescent substrate,<br />

native coelenterazine, and oxygen bound to the protein.<br />

Upon addition of excess calcium ions that fill the three<br />

calcium binding sites, the oxygen reacts with the substrate<br />

to yield a rapid flash of blue light centered at 485 nm. This<br />

emission reaches a peak within 100 msec and decays in<br />

less than 1 second.<br />

80,000<br />

40,000<br />

20,000<br />

0<br />

340 350 360 370 380 390 400 410 420 430 440<br />

Wavelength (nm)<br />

0<br />

470 480 490 500 510 520 530 540 550 560 570<br />

www.ArborAssays.com | info@ArborAssays.com | Ph: +1.734.677.1774<br />

25,000<br />

20,000<br />

15,000<br />

FLu<br />

10,000<br />

5,000<br />

Wavelength (nm)


FeAtures<br />

f Uses WB, IP, IF, ChIP, EIA, ELISA<br />

Abx <br />

monoclonal and polyclonal Antibodies<br />

DesCriptioN host uses siZe CAtALog No.<br />

epigeNetiCs<br />

Histone H3 Rabbit WB, IP, ChIP 100 µL A004-100UL<br />

DiMe-Lys4-Histone H3 Rabbit WB, IF 100 µL A006-100UL<br />

MonoMe-Lys4-Histone H3 Rabbit WB, IP, ChIP 100 µL A005-100UL<br />

TriMe-Lys4-Histone H3 Rabbit WB, IF, IP, ChIP 100 µL A007-100UL<br />

LSD1 Rabbit WB, IP 100 µL A003-100UL<br />

oxiDAtiVe stress<br />

L-Cysteine Mouse WB, IF, IHC, IP, ELISA 50 µg A002-50UG<br />

Glutathione Mouse WB, IF, IP, ELISA 50 µg A001-50UG<br />

Glutathione-DyLight® 488 Mouse WB, IF, FACS 50 µg A001F-50UG<br />

Glutathione-DyLight® 550 Mouse WB, IF, FACS 50 µg A001T-50UG<br />

pLAte CoAtiNg/DeteCtioN<br />

Mouse IgG, Fc, Aff. Pure Goat EIA, ELISA 10mg A008-10MG<br />

Mouse IgG, Fc, Aff. Pure Goat EIA, ELISA 25mg A008-25MG<br />

Rabbit IgG, Fc, Aff. Pure Goat EIA, ELISA 10mg A009-10MG<br />

Rabbit IgG, Fc, Aff. Pure Goat EIA, ELISA 25mg A009-25MG<br />

Sheep IgG, Aff. Pure Donkey EIA, ELISA 10mg A010-10MG<br />

Sheep IgG, Aff. Pure Donkey EIA, ELISA 25mg A010-25MG<br />

AFFiNity resiNs<br />

Cortisol Sheep Cortisol Isolation 500 µL R001-500UL<br />

WB=Western blotting: IP=Immunoprecipitation: IF=Immunofluorescence: IHC=Immunoohistochemistry:<br />

EIA=Enzyme <strong>immunoassay</strong>; ELISA=Enzyme-Linked ImmunoSorbant Assay: ChIP=Chromatin Immunoprecipitation<br />

typiCAL DAtA<br />

Immunofluorescence<br />

HeLa Cells stained with MxGSH monoclonal<br />

A001-50UG and goat anti-mouse IgG-FITC<br />

reLAteD proDuCts<br />

DyLight® is a registered trademark of Thermo Fisher Corp<br />

Glutathione Detection Kits Catalog No. K006-F1/F5/H1 Page 22<br />

Histone Demethylase Fluorescent Activity Kit Catalog Number K010-F1 Page 26<br />

www.ArborAssays.com | info@ArborAssays.com | Ph: +1.734.677.1774 43


44<br />

pdx <br />

enzyme inhibitors and Activators<br />

FeAtures<br />

f Unique, high quality Enzyme Inhibitors and Activators<br />

DesCriptioN uses siZe CAtALog No.<br />

DNA hypermethyLAtioN AgeNts<br />

5-Azacytidine DNA Methyltransferase Inh. 50/250 mg P012-50/250MG<br />

Decitibine DNA Methyltransferase Inh. 10/50 mg P015-10/50MG<br />

histoNe ACetyLtrANsFerAse iNhibitors<br />

C-646 p300/CBP HAT inhibitor 5/25 mg P014-5/25MG<br />

Garcinol Potent HAT inhibitor 5/25 mg P017-5/25MG<br />

hDAC iNhibitors<br />

Sodium Valproate HDAC Inhibitor 5 g P001-5G<br />

SAHA (Vorinostat) Catalytic HDAC Inhibitor 50/250 mg P004-50/250MG<br />

Phenyl Butyrate, Na Salt HDAC Inhibitor 1 G P007-1G<br />

Hexyl-4-pentynoic acid (HPA) Histone hyperacetylation 10/50 mg P008-10/50MG<br />

⇪ Arbor Assays exclusive!<br />

Trichostatin A Potent, Reversible Inhibitor 1 mg P010-1MG<br />

Apicidin Potent HDAC Inhibitor 1/5 mg P011-1/5MG<br />

BML-210 Potent HDAC inhibitor 5/25 mg P013-5/25MG<br />

histoNe methyLtrANsFerAse iNhibitors<br />

BIX-01294 G9a & GLP HMTase Inhibitor 5/25 mg P018-5/25MG<br />

NAD biosyNthesis iNhibitor<br />

FK-866, HCl Specific inhibitor of NAMPT 5/25 mg P006-5/25MG<br />

phosphoDiesterAse iNhibitor<br />

IBMX Pan-specific inhibitor of PDEs 100 mg/1G P019-100MG/1G<br />

sirt iNhibitors AND ACtiVAtors<br />

Sirtinol NAD-dependant HDAC Inh. 5/25 mg P016-5/25MG<br />

Resveratrol SIRT1 activator 100/500 mg P002-100/500MG<br />

BPDP (BML-278) Novel SIRT activator 5/25 mg P003-5/25MG<br />

Piceatannol Activator of SIRT1 5/25 mg P009-5/25MG<br />

EX-527 Selective SIRT1 inhibitor 5/25 mg P005-5/25MG<br />

histoNe DemethyLAse iNhibitors<br />

Tranylcypromine LSD1 Inhibitor 10 mM X042-1EA<br />

For structures please see page 45.<br />

www.ArborAssays.com | info@ArborAssays.com | Ph: +1.734.677.1774


pdx<br />

enzyme Activator and inhibitor structures<br />

5-Azacytidine Apicidin BIX-01294 BML-210<br />

Catalog No. P012 Catalog No. P011 Catalog No. P018 Catalog No. P013<br />

BML-278 C-646 Decitibine EX-527<br />

Catalog No. P003 Catalog No. P014 Catalog No. P015 Catalog No. P005<br />

FK-866 Garcinol HPA IBMX<br />

Catalog No. P006 Catalog No. P017 Catalog No. P008 Catalog No. P019<br />

Phenylbutyrate Piceatannol Resveratrol SAHA<br />

Catalog No. P007 Catalog No. P009 Catalog No. P002 Catalog No. P004<br />

Sirtinol Trichostatin A Tranylcypromine Valproate<br />

Catalog No. P016 Catalog No. P010 Catalog No. X042-1EA Catalog No. P001<br />

www.ArborAssays.com | info@ArborAssays.com | Ph: +1.734.677.1774 45


46<br />

technical information<br />

teChNiCAL iNDex<br />

f Immunoassay Information<br />

f<br />

f<br />

f<br />

f<br />

Getting the Most From Your Assay<br />

Plate Reader Set Up<br />

Sample Extraction<br />

Premium Rules of Assays<br />

immuNoAssAy iNFormAtioN<br />

The basic components of any <strong>immunoassay</strong> system are three-fold; an antigen or antibody that we would like to detect<br />

and quantitate, specific antibodies to this antigen, and a system to measure the amount of the antigen in a given sample.<br />

In many cases a number of other assays materials are necessary to allow for quick and convenient measurement. This<br />

simplified system has been used in several different ways and the three most common are outlined below. NOTE: “ELISA”<br />

has been mistakenly used for all 3 types (and more) of the systems described below. We will interchange EIA and ELISA<br />

in marketing materials but, to us, an EIA is a small molecule <strong>immunoassay</strong>, a Sandwich assay is for larger proteins and<br />

biomolecules, and a “real” ELISA is an assay to measure antibodies.<br />

1. <strong>eiA</strong><br />

This competitive assay system is best exemplified by a typical EIA system. Here a single antibody to a small molecular<br />

weight antigen, typically less than 10,000 Dalton, is used. This antibody, at a very specific defined and limited concentration,<br />

competitively binds the antigen in the sample and the antigen labeled with some detection system, like Horseradish<br />

Peroxidase. The amount of the bound antigen added to the reaction is measured at the end of the immunological binding<br />

reaction and the percentage bound is inversely proportional to the amount of unlabeled antigen. Separation at the end of the<br />

immunological binding reaction can be done by a number of systems but typical separation systems use a microtiter plate, or<br />

beads. This type of reaction, along with its variations, is the only method possible for small molecular weight antigens, such<br />

as steroids, drugs, lipids, and peptides.<br />

The typical format for one of our EIAs is shown below. We use peroxidase as the detection enzyme because of its stability,<br />

turnover number and lack of interferences. We separate the antibody-bound labeled antigen at the end of the immunological<br />

binding reaction using a secondary antibody coated microtiter plate that specifically binds the primary antibody. The excess<br />

reagents are washed out of the plate. The resulting signal from the plate bound labeled antigen is inversely proportional to<br />

the amount of antigen in the sample.<br />

Sample Antigen<br />

Primary<br />

Antibody<br />

Capture Antibody<br />

Antigen-Peroxidase Conjugate<br />

see page 15 for typical binding Curves.<br />

Colorimetric Chemiluminescent<br />

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technical information Continued<br />

2. sandwich Assays<br />

Also called immunometric assays, they use 2 or more specific antibodies to “sandwich” the antigen. Typically one antibody<br />

is bound to the separation system, such as a microtiter plate, and one antibody is used to detect the antigen. It can be seen<br />

that the antigen is captured between the 2 antibodies, one attached to the solid phase, the other labeled with an enzyme.<br />

Typically the amount of solid phase antibody and enzyme-conjugated antibody are in a large excess over the amount of<br />

antigen in the sample. This makes the kinetics of binding of the antigen to the solid phase, and the antibody conjugate to<br />

the antigen to be pseudo-first order, resulting in very rapid kinetics and high sensitivity. The result is an assay that produces<br />

a signal that is proportional to the amount of antigen in solution.<br />

Sample Antigen<br />

Primary Capture Antibody<br />

Primary Antibody-Peroxidase Conjugate<br />

Colorimetric Chemiluminescent<br />

3. eLisA Assays<br />

The real “ELISA” format uses a solid phase coated with either a specific protein, killed or neutralized virus, or synthetic<br />

peptide fragments. It is commonly used to detect the presence of antibodies to specific antigenic sites on proteins and<br />

viruses. These assays have been used extensively in AIDS and hepatitis testing. Samples, typically from donated blood, are<br />

applied to the solid phase. Any antibodies to the virus, suggesting viral exposure, will bind to the viral antigen on the solid<br />

phase. After a wash step, a second antibody, labeled with an enzyme is added. This second antibody is specific to human<br />

IgG or IgM and binds to the sample antibody bound to the solid phase antigen.<br />

4. CLiA or ChemiLuminescent immunoAssays<br />

This is an assay where the readout is the emission of light from a CLIA substrate is substituted for the TMB and the reaction<br />

of the captured antigen molecule labeled with peroxidase, or the peroxidase conjugated detection antibody. CLIAs<br />

use white plates and these white plates are coated using identical conditions as for the clear plates used in EIAs. In our<br />

peroxidase based readout CLIA assays after the final wash step the chemiluminescent substrate is added to the plate and<br />

the intensity is read immediately. We specify 0.1 second read time per well using our CLIA substrate. This allows a typical<br />

96-well plate to be read in about 10 seconds.<br />

5. Antibody generation<br />

Antigen purity<br />

In order to raise an antibody to a particular antigen the first requirement is the need for a reliable supply of the pure high<br />

quality antigen. This is very important. If the antigen injected into an animal is impure the resulting antisera may recognize<br />

the impurities with equal or greater affinity thereby invalidating the whole measurement method. The purity of the antigen<br />

is therefore of the utmost importance.<br />

Attachment Chemistry<br />

If you wish to raise an antibody to a small molecular weight antigen, such as one of the prostaglandins we work with, the<br />

molecule is not large enough to elicit an antigenic response by itself. For molecules up to about 5,000-10,000 molecular<br />

weight the molecule should be covalently conjugated to a carrier protein for optimum antigenic recognition. The injected<br />

antigen conjugated protein, will give rise to a range of antibodies, some recognizing the carrier protein and some the antigen<br />

conjugated to the protein. Suitable proteins are albumins, such as BSA and ovalbumin, or keyhole limpet hemocyanin<br />

(KLH). The antigen must be attached at a high ratio to the protein. If the antigen is larger than 5,000 to 10,000 molecular<br />

weight then it can be used directly without any chemical modification.<br />

www.ArborAssays.com | info@ArborAssays.com | Ph: +1.734.677.1774 47


48<br />

technical information Continued<br />

To attach a small molecular weight compound to a carrier protein the typical methods employed use any reactive groups<br />

on the antigen. For example, with prostaglandins we activate the carboxylic acid group on the prostaglandin molecule<br />

before conjugating to form an amide bound with the lysine groups on the particular carrier protein. Suitable conjugation<br />

methods can be found in the books by T. Chard, in “An Introduction to Radio<strong>immunoassay</strong> & Related Techniques”, (1990),<br />

4th Ed., Elsevier, Amsterdam and by P. Tijssen, in “Practice & Theory of Enzyme Immunoassays”, (1985), Elsevier, Amsterdam.<br />

Both books have methods on conjugating small molecules to carrier proteins. After conjugation the labeled carrier<br />

protein is purified and aliquoted.<br />

Animal Facility and host<br />

The next choices that have to be made are the selection of the facility for raising the antisera and the species of animal to<br />

be used. Avoid companies that raise their own antisera for sale; they may be “tempted” to sell your antisera. A convenient<br />

way to avoid this is to use only an identifier, e.g., “Arbor123”.<br />

Once you have selected a suitable vendor then the selection of host animal should be chosen. Most polyclonals are raised<br />

in rabbits and monoclonals in mice. The choice of host can be very wide depending on both volume of antisera needed<br />

and any expected host interferences. For most experiments rabbits will provide an abundant quantity of antisera, however,<br />

for large scale applications, such as microtiter plate coating, goats, sheep, donkeys, horses, or the use of monoclonals will<br />

give you an “unlimited” supply. Note that raising monoclonals is about 3 times more expensive than polyclonals and the<br />

antisera obtained may not have as high an affinity or may not be as stable as a polyclonal source.<br />

Antisera evaluation<br />

The evaluation of the antisera produced is typically a major task. It will involve assessment of affinity or sensitivity, cross<br />

reactivity, and stability among other parameters. When we develop an assay using a newly obtained antibody we will<br />

evaluate each individual antisera in that order, determining if the antisera has a high enough affinity to allow detection of<br />

the antigen in the normal range. This ranking has allowed us to produce a number of <strong>immunoassay</strong>s with superior performance<br />

characteristics.<br />

Having produced a sufficient quantity of antisera with good cross reactivity and sensitivity, the evaluation process should<br />

then be followed by stability and other parameters. In general polyclonal antisera are very stable, unstable IgG molecules<br />

have probably been eliminated within the host prior to the sera being isolated. In our experience polyclonal antisera are<br />

stable for over 1 year at 4°C in suitable buffers containing carrier proteins and preservatives. However, evaluate the stability<br />

of each antisera for yourself.<br />

Monoclonal antibodies can have different properties due to the fact that they are a single IgG or IgM species. In general<br />

IgG1 species are the most stable although other species may display adequate stability. The monoclonals may also be IgM<br />

and require special handling. Obviously each monoclonal is a unique molecule and the greatest care and attention should<br />

be used for the storage of any clones. The clones should be stored in multiple locations so that a catastrophe at any one<br />

site does not destroy the ability to produce more antibody. Each storage location should have emergency backup and<br />

alarm features. In our hands monoclonal antibodies of the IgG 1 species are also stable in solution at 4°C in buffers containing<br />

carrier proteins and preservatives.<br />

gettiNg the most From your AssAy<br />

As Albert Einstein once said “If we knew what we were doing, it wouldn’t be called Research”. This applies to your experiment<br />

as well. If you knew that the concentration of “X” was Y pg/mL you probably would not have bought our <strong>kit</strong> for<br />

measuring “X”. So before you use the whole <strong>kit</strong> we suggest you try the following:<br />

1.<br />

2.<br />

3.<br />

Evaluate the dilution needed to measure the analyte by taking a normal sample, and make serial 1:10 dilutions with the<br />

<strong>kit</strong> assay buffer or diluent.<br />

Run a single well of the serial 1:10, 1:100, etc dilutions in the <strong>kit</strong> to determine the dilution necessary for accurate analyte<br />

measurement.<br />

Depending on if you expect the concentrations in your other samples will be higher or lower than this normal sample,<br />

determine the dilution where the signal starts to change in a linear or predictable fashion.<br />

www.ArborAssays.com | info@ArborAssays.com | Ph: +1.734.677.1774


4.<br />

5.<br />

technical information Continued<br />

Dilute the samples aiming for them to be at the appropriate part of the curve for your experiment.<br />

If you are performing a fluorescent or chemiluminescent assay you can also ensure that the reader is performing as<br />

expected with the high and low signal giving you acceptable signals.<br />

pLAte reADer set up<br />

The proper experimental set up is very important to get the most from our <strong>kit</strong>s. For most CL experiments the set up is easy.<br />

CL is read without any filters or wavelength selection and so the only parameter that needs to be addressed is the Gain<br />

setting. Some instruments will allow you to alter the gain and you need to set the lowest signal well to be close to zero RLU<br />

or FLU. Your highest signal well should be set at about 90% of the dynamic range of the reader. These settings will give you<br />

the best signal-to-noise (S/N) ratio.<br />

FL assays require the same set up as CL measurements above but with the added step of ensuring the excitation (Ex) and<br />

emission (Em) settings are appropriate. FL readers may be either filter or monochromator readers. Filter readers require that<br />

the Ex amd Em filters are appropriate for the <strong>kit</strong>. Please contact us if you have questions. Monochromator readers allow the<br />

precise wavelength to be set.<br />

If you have any other questions about gain or filter selections please contact your plate reader manufacturer.<br />

sAmpLe extrACtioN<br />

Visit our website for extraction protocols at: http://www.arborassays.com/resources/lit.asp There are typical protocols for<br />

a variety of different sample types. Below are outlines of 3 helpful protocols for different samples.<br />

Cyclic Nucleotides in tissues. Grind a weighed amount of frozen tissue in a homogenizer in the <strong>kit</strong> Sample Diluent. Use<br />

the Sample Diluent supernatant for the cAMP or cGMP assay, and if you require a protein determination, use a BCA protein<br />

type assay to determine protein concentration. See the <strong>kit</strong> manuals for more information.<br />

prostaglandins from tissues. Grind a weighed amount of frozen tissue in a homogenizer in pH 4 0.1M phosphate<br />

buffer:absolute ethanol 30:70 using 10-50 µL per mg of tissue. Centrifuge and speed evaporate the supernatant until dry.<br />

Reconstitute in the <strong>kit</strong> Assay Buffer. See the <strong>kit</strong> manuals for more information.<br />

Fecal steroids in Field situations. Take a weighed amount of fecal material and add methanol:water 80:20 using a ratio of<br />

extraction media to wet feces of 2.5:1. Samples can be stored in this matrix if kept at -20°C. When processing samples for<br />

assay purposes, take the thawed sample and shake for 20 minutes. Centrifuge and collect the supernatant. These samples<br />

can be kept at -20°C for later processing; diluted at least 1:50 in <strong>kit</strong> assay buffer for assay immediately; or speed vacuum<br />

evaporated to dryness. See the <strong>kit</strong> manuals for more information.<br />

premium ruLes oF AssAys:<br />

1. sample matrix. Your samples must be in the identical matrix as the standards to obtain a valid reading.<br />

2. Data Analysis. We suggest that you always use the 4PLC software that is available for almost all plate readers. For<br />

small molecule assays, such as cortisol, cAMP or PGE , a 4PLC program must be used. Most programs have provisions<br />

2<br />

to flag out of range values and to give you CVs for both the signal and the concentration.<br />

3. Contact us before you run the Assay. If you are in any doubt about the protocol, how to treat your samples, how to<br />

handle any of the reagents, etc. please contact us by E mail at Technical@ArborAssays.com or call us at 734-677-1774<br />

before you start the assay. We will speedily answer your question and make suggestions so you can get the most out<br />

of your experiment.<br />

www.ArborAssays.com | info@ArborAssays.com | Ph: +1.734.677.1774 49


© 2012<br />

DetectX®, ThioStar®, Arbor Assays® Logo are<br />

Registered Trademarks of Arbor Assays LLC

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