Anti-Tyrosine Hydroxylase, clone 2/40/15 - Millipore

Anti-Tyrosine Hydroxylase, clone
2/40/15
Monoclonal Antibody
Cat. # MAB5280
Lot # LV1478876
pack size: 40 µg
Store at 2-8ºC
FOR RESEARCH USE ONLY
NOT FOR USE IN HUMANS
Certificate of Analysis
page 1 of 2
Species
Antibody Epitope/
Host
Molecular
Applications
Accession #
Cross-Reactivity Isotype
Region
Species
Weight
WB, IH H, B, Ch, M, Qu, R IgG2a N/A M 56-60 kDa NM_000360.3
Background
Tyrosine hydroxylase plays an important role
in the physiology of adrenergic neurons. It is
the first and rate-limiting enzyme involved in
the biosynthesis of the catecholamines
Dopamine and Norepinephrine from tyrosine.
TH is, therefore, a useful marker for
dopaminergic and noradrenergic neurons. The
enzymatic activity of TH requires ferrous ions
as cofactors and is believed to be regulated by
phosphorylation. At least four isoforms of
human TH have been identified which result
from alternative splicing.
Presentation
Purified mouse monoclonal IgG2a liquid in
buffer containing 0.02 M Phosphate buffer,
0.25 M NaCl, pH 7.6 with 0.1% sodium azide.
Concentration
1 mg/mL
Specificity
Tyrosine hydroxylase (TH) catalyzes the ratedetermining
initial step in the biosynthesis of
catecholamines such as dopamine,
noradrenaline and adrenaline. Therefore the
antibody is used to stain dopaminergeous and
adrenergeous neurons as well as the
chromaffine tissue of the adrenal medulla. Cell
lines such as the PC 12 line, which is derived
from a rat pheochromocytoma also, show
positive reactions. In immunoblots of adrenal
medulla tissue, the antibody recognizes a
single protein band in the molecular weight
range of 56 - 60 kD.
Immunogen
Purified tyrosine hydroxylase (EC 1.14.16.2)
from a rat pheochromocytoma (2).
Molecular Weight
56-60 kDa
Method of Purification
Protein A Purified
Storage and Handling
Stable for 6 months at 2-8ºC in undiluted
aliquots from date of receipt.
Control
Positive Control: Brain (sympathetic nerve
terminals) and adrenal gland tissue
Quality Control Testing
Routinely evaluated by Western Blot on PC12
lysates.
Western Blot Analysis: 1:1000 dilution of this
lot detected Tyrosine Hydroxylase on 10 ug of
PC12 lysates.
Western Blot Analysis:
PC12 lysate was resolved by
electrophoresis, transferred to
PVDF membrane and probed with
anti-Tyrosine Hydroxylase (1:1000
dilution of this lot).
Proteins were visualized using a
goat anti-mouse secondary
antibody conjugated to HRP and a
chemiluminescence detection
system.
Arrow indicates protein Tyrosine
Hydroxylase (~64 kDa).
References
1. Nafia, Imane, et al. (2008). J. Neurochem. 105:
484-96.
2. Casarejos, M J, et al. (2005). J Neurochem.
94:1005-14.
3. Serrano, A., et al. (2005). Neuropharmacology.
49:208-219.
4. Gauda, E.B., et al. (2004). J. Applied
Physiology. 96:384-391.
5. de Bernardo, S., et al. (2004). J.
Neurochemistry. 91:667-682.
APPLICATION LEGEND: WB Western Blotting IP Immunoprecipitation IC Immunocytochemistry IF Immunofluorescence
IH Immunohistochemistry (Tissue)
SPECIES LEGEND: B Bovine Ch Chicken H Human M Mouse Qu Quail R Rat Rb Rabbit WR Most Common Vertebrates
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Rev.A/2008-07-17/MAB5280CA/JM

Anti-Tyrosine Hydroxylase, clone 2/40/15
Cat # MAB5280
Lot # LV1478876
Additional Research Applications
page 2 of 2
Immunohistochemistry: Representative image from a previous lot.
A previous lot of this antibody was used in immunohistochemistry.
Optimal working dilutions must be determined by end user.
PROTOCOL
Western Blot
1. Perform SDS-polyacrylamide gel electrophoresis (SDS-PAGE) on cell lysate and transfer the proteins to a PVDF membrane. Wash the PVDF
membrane twice with water.
2. Block the blotted PVDF membrane in freshly prepared 5% BSA with 0.05% Tween ® -20 for 1 hour at room temperature with constant agitation.
3. Incubate the PVDF with the recommended dilution of Anti-Tyrosine Hydroxylase, clone 2/40/15 diluted in freshly prepared 5% BSA for 1 hour
at room temperature or overnight with agitation at 2-8°C.
4. Wash the PVDF 3 times with TBST.
5. Incubate the PVDF in the secondary reagent of choice (a goat anti-mouse HRP conjugated IgG, Catalog # 12-349 1:1000 dilution was used) in
5% milk for 1 hour with agitation at room temperature.
6. Wash the PVDF 3-5 times with TBST.
7. Use Spray and Glow Catalog # 17-373 to visualize results. Use as directed.
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