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1. Martinon F,. et al., 2002. The inflammasome: a molecular platform triggering activation of inflammatory caspases and processing of proIL-beta. Mol Cell. 2002 10(2):417-26. 2. Miao EA. et al., 2006. Cytoplasmic flagellin activates caspase-1 and secretion of interleukin 1beta via Ipaf. Nat Immunol. 7(6):569-75. 3. Boyden ED & Dietrich WF., 2006. Nalp1b controls mouse macrophage susceptibility to anthrax lethal toxin. Nat Genet. 38(2):240-4. 4. Mariathasan S & Monack DM., 2007. Inflammasome adaptors and sensors: intracellular regulators of infection and inflammation. Nat Rev Immunol. 7(1):31-40. 5. Martinon F, & Tschopp J., 2004. Inflammatory caspases: linking an intracellular innate immune system to autoinflammatory diseases. Cell. 117(5):561-74. 6. Pelegrin P, & Surprenant A., 2007. Pannexin-1 couples to maitotoxin- and nigericin-induced interleukin-1beta release through a dye uptake-independent pathway. J Biol Chem. 282(4):2386-94. 7. Solle M. et al., 2001. Altered cytokine production in mice lacking P2X(7) receptors. J Biol Chem. 276(1):125-32. 8. Kanneganti TD, et al., 2007. Pannexin-1-mediated recognition of bacterial molecules activates the cryopyrin inflammasome independent of Toll-like receptor signaling. Immunity. 26(4):433-43. 9. Martinon F. et al., 2006. Gout-associated uric acid crystals activate the www.invivogen.com/inflammasome NALP3 inflammasome. Nature. 440(7081):237-41. 10. Dostert C. et al., 2008. Innate immune activation through Nalp3 inflammasome sensing of asbestos and silica. Science. 320(5876):674-7. 11. Cassel SL. et al., 2008. The Nalp3 inflammasome is essential for the development of silicosis. Proc Natl Acad Sci U S A. 105(26):9035-40. 12. Hornung V. et al., 2008. Silica crystals and aluminum salts activate the NALP3 inflammasome through phagosomal destabilization. Nat Immunol. 9(8):847-56. 13. Eisenbarth SC. et al., 2008. Crucial role for the Nalp3 inflammasome in the immunostimulatory properties of aluminium adjuvants. Nature. 453(7198):1122-6. 14. Li H. et al., 2008. Cutting edge: inflammasome activation by alum and alum's adjuvant effect are mediated by NLRP3. J Immunol. 181(1):17- 21. 15. Hornung V. et al., 2009. AIM2 recognizes cytosolic dsDNA and forms a caspase-1activating inflammasome with ASC. Nature. 458(7237):514-8. 16. Fernandes-Alnemri T. et al., 2009. AIM2 activates the inflammasome and cell death in response to cytoplasmic DNA. Nature. 458(7237):509-13. 17. Bürckstümmer T. et al., 2009. An orthogonal proteomicgenomic screen identifies AIM2 as a cytoplasmic DNA sensor for the inflammasome. Nat Immunol. 10(3):266-72. 97 INNATE IMMUNITY 3
INNATE IMMUNITY 3 Inflammasome Inducers & Inhibitors The inflammasome is a multi-protein complex involved in the production of mature IL-1b. It is activated by a large variety of microbial molecules, danger signals and crystalline substances. InvivoGen provides a selection of these molecules known to induce the assembly of the NLRP3 or AIM2 inflammasomes. We also offer some inhibitors to confirm the ability of a given molecule to activate the inflammasome or to test the importance of phagocytosis or potassium efflux in this process. These inflammasome inducers and inhibitors have been validated using the THP-1/HEK-Blue -IL-1b Assay (see page 100). 98 PRODUCT DESCRIPTION Inflammasome Inducers Alum Crystals Aluminium potassium sulfate 10 - 200 μg/ml 1 g tlrl-alk ATP Adenosine 5′-triphosphate disodium salt 5 mM 1 g tlrl-atp CPPD Crystals Calcium pyrophosphate dihydrate 50 - 200 μg/ml 5 mg tlrl-cppd Hemozoin NEW Synthetic heme crystal 50 - 400 μg/ml 5 mg tlrl-hz MSU Crystals Monosodium urate (uric acid) 50 - 200 μg/ml 5 mg tlrl-msu Nigericin Nigericin, sodium salt 1 μM 10 mg tlrl-nig Poly(dA:dT), double-stranded B DNA Poly(dA-dT)•poly(dT-dA)/LyoVec complex 1 - 5 μg/ml 100 μg tlrl-pat Inflammasome Inhibitors Inflammasome Inducers Alum Crystals - NLRP3 Inflammasome Inducer Aluminum hydroxide and potassium salts (alum) are commonly used vaccine adjuvants. Adjuvants are vaccine additives that stimulate the immune system without having any specific antigenic effect. Alum has been demonstrated to activate caspase-1 and triggers IL-1b and IL-18 secretion 1 . All alum preparations contain crystals. The alum-induced release of IL-1b in macrophages is dependent on NLRP3 and ASC, indicating that alum triggers inflammation through activation of the NLRP3 inflammasome 2 . Alum has been shown to trigger NLRP3 activation through lysosomal destabilization 2 . ATP - NLRP3 Inflammasome Inducer Adenosine triphosphate (ATP), a potassium efflux agent, can trigger the activation of NLRP3 inflammasome in response to PAMPs, such as LPS and peptidoglycan. It stimulates the caspase-1-dependent cleavage and secretion of IL-1b from LPS-stimulated cells 3 . ATP triggers the opening of the nonselective cation channel of the purinergic P2X7 receptor, followed by the gradual opening of a larger pore. The larger pore is attributed to pannexin-1, which is recruited upon P2X7 receptor activation 4 . Activation of the P2X7 receptor results in potassium efflux which is necessary for activation of the posttranslational maturation of IL-1b 5 . Hemozoin - NLRP3 Inflammasome Inducer NEW Hemozoin is a dark-brown heme crystal produced by the intraerythrocytic parasite Plasmodium, the causative agent of malaria. Hemozoin is taken up by macrophages initiating signals that lead to the production of IL-1b. Hemozoin-induced IL-1b production is dependent on the activation of the NLRP3 inflammasome 6, 7 . Synthetic hemozoin has been shown to possess adjuvant properties that differ depending on the method of synthesis 8 . InvivoGen provides a chemically synthesized hemozoin using an acidic method. MSU & CPPD Crystals - NLRP3 Inflammasome Inducers Crystals of monosodium urate (MSU) and calcium pyrophosphate dihydrate (CPPD) are the aetiological agents of the inflammatory joint diseases gout and pseudo-gout, respectively. Both pathogenic crystals have been recently shown to be potent activators of caspase-1 through the NLRP3 inflammasome 9 . Involvement of the inflammasome is suggested by the finding that macrophages from mice deficient in various components of the inflammasome are defective in crystal-induced IL-1b induction. NLRP3 activation requires the phagocytosis of crystals that leads to lysosomal damage which appears to be the signal recognized by the inflammasome resulting in its activation 2 . www.invivogen.com/inflammasome WORKING CONCENTRATION QUANTITY CATALOG CODE Glybenclamide Proton pump inhibitor 25 μg/ml 1 g tlrl-gly Z-VAD-FMK Pan-caspase Inhibitor 10 μg/ml (20 μM) 1 mg tlrl-vad
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INNOVATION WITHIN REACH InvivoGen
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CONTENTS AT A GLANCE 1 CELL CULTURE
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4 CYTOKINE SIGNALING TABLE of CONTE
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Mycoplasma Detection & Elimination
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PlasmoTest Contents - HEK-Blue -2
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Primocin - The Antimicrobial Shiel
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SEAP Reporter Gene System Secreted
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HEK-Blue Detection - Real-Time Det
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Blasticidin S Description Blasticid
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LyoVec New formulation LyoVec is
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Poly-Arginine-HA Tagged Reprogrammi
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LENTI-Smart - Lentiviral Vector Pr
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Lentiviral Vector Production and Ce
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Antibody generation using pFUSE-CHI
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Engineering Fc regions for altered
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pBROAD & pWHERE - Optimized Vector
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pSELECT-Tag - Expression of a GFP-,
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pVIVO - Expression of Two Genes of
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pORF Description pORF plasmids feat
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Prom A-List - Human and Rodent Pro
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PROMOTER GENE TISSUE DISTRIBUTION S
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ChemiComp E. coli ChemiComp E. coli
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Fast-Media ® Preparation 1. Empty
Page 48 and 49: RIG-I-like Receptors 54 RLR & RLR-R
Page 50 and 51: Toll-Like Receptors Toll-Like Recep
Page 52 and 53: www.invivogen.com/innate-immunity 5
Page 54 and 55: IPAF and NAIP5 IPAF (NLRC4, CLAN/CA
Page 56 and 57: C-Type Lectin Receptors C-type lect
Page 58 and 59: pUNO pUNO-HA Toll-Like Receptors (T
Page 60 and 61: GENE NAME/ALIASES DESCRIPTION CAT.
Page 62 and 63: pDUO - Gene Associations Features a
Page 64 and 65: pZERO-TLR & pDeNy - Dominant Negati
Page 66 and 67: 293/TLR & 293/NOD Clones 293/TLR Cl
Page 68 and 69: Response of HEK-Blue hTLR2 cells t
Page 70 and 71: Ramos-Blue Cells - NF-kB/AP-1 Repo
Page 72 and 73: MEF Cells C3H/TLR4mut & C3H/WT MEFs
Page 74 and 75: Antibodies for Detection ANTIBODY D
Page 76 and 77: - NF-kB binding site: Nuclear facto
Page 78 and 79: PRODUCT ORIGIN/DESCRIPTION TLR2 Ago
Page 80 and 81: PRODUCT ORIGIN/DESCRIPTION TLR9 Ago
Page 82 and 83: PRODUCT ORIGIN/DESCRIPTION Dectin-1
Page 84 and 85: LPS-PG (Porphyromonas gingivalis) R
Page 86 and 87: • FLA-ST ultrapure was purified b
Page 88 and 89: 1. Zhao H. et al., 2004. Contributi
Page 90 and 91: 1. Hornung V. et al., 2006. 5'-Trip
Page 92 and 93: TLR/NOD Ligand Screening InvivoGen
Page 94 and 95: Signal Transduction Inhibitors Invi
Page 96 and 97: Pepinh-MYD - MyD88 inhibitory pepti
Page 100 and 101: Nigericin - NLRP3 Inflammasome Indu
Page 102 and 103: Autophagy is a pathway by which cyt
Page 104 and 105: 4 CYTOKINE SIGNALING Cytokine Genes
Page 106 and 107: Blue Cytokine Reporter Cells Blue
Page 108 and 109: HEK-Blue IFN-a/b Cells - Human Typ
Page 110 and 111: HEK-Blue IL-6 Cells - IL6 Reporter
Page 112 and 113: HEK-Blue IL-1b Cells - IL-1b Repor
Page 114 and 115: HEK-Blue IL-33/IL-1b Cells - IL-33
Page 116 and 117: HEK-Blue CD40L Cells - CD40L Repor
Page 118 and 119: 5 ANTIBODIES & VACCINATION Antibodi
Page 120 and 121: Antibody generation using pFUSE-CHI
Page 122 and 123: Immunoglobulin A The mucosal surfac
Page 124 and 125: IgA Detection and Quantification In
Page 126 and 127: IgG Antibodies InvivoGen offers a s
Page 128 and 129: Mediators? 1. Leroux-Roels G., 2010
Page 130 and 131: Poly(I:C) Synthetic double-stranded
Page 132 and 133: pVAC - Induction of Neutralizing An
Page 134 and 135: CpG-Free DNA CpGs and the Immune Re
Page 136 and 137: pCpGfree-basic & pCpGfree-promoter
Page 138 and 139: pCpGfree-siRNA & pCpGfree-siRNADUO
Page 140 and 141: Custom-Made CpG-Free Genes InvivoGe
Page 143 and 144: RNA INTERFERENCE 7 psiRNA System I
Page 145 and 146: RNA INTERFERENCE 7 psiRNA-h7SK - 7S
Page 147 and 148: RNA INTERFERENCE 7 psiTEST System
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RNA INTERFERENCE 7 Ready-Made psiRN
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RNA INTERFERENCE 7 Custom-Made psiR
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INHIBITORS 8 Inhibitors PRODUCT DES
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INHIBITORS 8 Pattern Recognition Re
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INHIBITORS 8 Triptolide - NF-kB Inh
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INHIBITORS 8 Heat Shock Protein Inh
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9 CUSTOM SERVICES TLR/NOD Ligand Sc
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CUSTOM SERVICES 9 Custom Cloning :
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CUSTOM SERVICES 9 Prices 164 TERMS
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166 PRODUCT (QUANTITY) CAT. CODE PA
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A 17-AAG 17-AAGH2 17-AEP-GA 2-Amino
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- ODN 4084-F - ODN INH-1 - ODN INH-
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ARGENTINA Genbiotech S.R.L. Tel: +5
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