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Signal Transduction Inhibitors InvivoGen offers a selection of known inhibitors of the signaling pathways initiated by PRRs. These inhibitory molecules intervene in the different steps of PRR activation and signaling cascade, including ligand binding, interaction with adapters and activation of MAP kinases and NF-kB. PRODUCT DESCRIPTION ENDOTOXIN LEVELS WORKING CONCENTRATION QUANTITY CATALOG CODE 2-Aminopurine PKR inhibitor
INNATE IMMUNITY 3 AG490 - JAK2 Inhibitor AG490 is a specific and potent inhibitor of the Janus kinase 2 protein (JAK2) 1 . JAK2 regulates the phosphorylation of JNK, primarily through PI3K. It has been established that JAK2 plays an important role in TLR-mediated biological responses, blocking TLR4-mediated responses to LPS 2 and TLR5-mediated responses to flagellin 3 . 2-Aminopurine - PKR Inhibitor 2-aminopurine (2-AP) is a potent inhibitor of double-stranded RNA (dsRNA)-activated protein kinase (PKR), a critical mediator of apoptosis. PKR is phosphorylated and activated by dsRNA and poly(I:C) and contributes to the induction of type I interferons, such as IFN-b, which can further increase its expression 4 . PKR plays also a role in TLR-induced antiviral activities as an intermediary in TLR3, TLR4 and TLR9 signaling 5 . . Bay 11-7082 - IkB-a Inhibitor Bay 11-7082 is an irreversible inhibitor of TNF-a-induced IkB-a phosphorylation resulting in the inactivation of NF-kB 6 . TNF-a-dependent effects of NF-kB are important for TLR expression and cytokine production 7 . BX795 - TBK/IKKε Inhibitor BX795, an aminopyrimidine compound, was developed as an inhibitor of 3-phosphoinositide-dependent kinase 1 (PDK1) 8 . It was recently shown to be a potent inhibitor of the IKK-related kinases, TANK-binding kinase 1 (TBK1) and IKKε, and hence of IRF3 activation and IFN-β production 9 . BX795 inhibits the catalytic activity of TBK1/IKKε by blocking their phosphorylation. Celastrol - NF-kB Inhibitor Celastrol is a triterpenoid compound isolated from the medicinal plant Tripterygium wilfordii known for its anti-inflammatory properties. Its mode of action and spectrum of cellular targets are still poorly understood. Celastrol was recently shown to act as an effective inhibitor of the transcription factor NF-kB resulting in the attenuation of nitric oxide and proinflammatory cytokine production 10 . Chloroquine - Inhibitor of Endosomal Acidification Chloroquine is a lysosomotropic agent that prevents endosomal acidification 11 . It accumulates inside the acidic parts of the cell, including endosomes and lysosomes. This accumulation leads to inhibition of lysosomal enzymes that require an acidic pH, and prevents fusion of endosomes and lysosomes. Chloroquine is commonly used to study the role of endosomal acidification in cellular processes, such as the signaling of intracellular TLRs 12 . CLI095 - TLR4 Signaling Inhibitor CLI095, also known as TAK-242, is a novel cyclohexene derivative that suppresses specifically TLR4 signaling, inhibiting the production of NO and pro-inflammatory cytokines 13 . It acts by blocking the signaling mediated by the intracellular domain of TLR4, but not the extracellular domain. It potently suppresses both ligand-dependent and -independent signaling of TLR4 14 . Cyclosporin A - Calcineurin inhibitor NEW Cyclopsporin A, a calcineurin inhibitor, exerts its immunosuppressive effects through the down-regulation of NFAT (nuclear factor of activated T cells), thus preventing the transcription of T cell effector cytokines. NFAT has been implicated in the downstream signaling of Dectin-1 15 . Conversely, it has been demonstrated that the inhibition of calcineurin in macrophages can trigger TLR signaling and enhance NF-κB activation 16 . Gefitinib - RIP2 Tyrosine Kinase inhibitor NEW Gefitinib (also known as IRESSA) is a selective inhibitor of epidermal growth factor (EGFR), a growth factor that plays a pivotal role in the control of cell growth, apoptosis, and angiogenesis. Recent studies demonstrated that Gefitinib can inhibit NOD2-induced cytokine release and NF-kB activation by inhibiting RIP2 (receptor-interacting protein 2) tyrosine phosphylation which is critical for activation of NOD2 downsream signaling pathways 17 . H-89 - PKA Inhibitor H-89 is a selective, potent cell permeable inhibitor of cAMP-dependent protein kinase (PKA) 18 . It can be used to determine the role of PKA in TLR and other PRR mediated signaling. PKA has be shown to participate in the TLR-mediated TREM-1 expression on macrophages following LPS stimulation 19 . Leptomycin B - Nuclear export inhibitor NEW Leptomycin B, an inhibitor of nuclear export, can be used to study nucleo-cytoplasmic translocation. It has been demonstrated that Leptomycin B can provoke the nuclear accumulation of proteins that shuttle between the cytosol and nucleus such as MAPK, MAPKAP kinase 2, IRAK-1 and NLRC5 20 . The cellular target of Leptomycin B has been identified as CRM1 (exportin 1), an evolutionarily conserved receptor for the nuclear export signal of proteins. LY294002 - PI3K Inhibitor LY294002 is a potent, cell permeable inhibitor of phosphatidylinositol 3-kinase (PI3K) that acts on the ATP binding site of the enzyme 21 . The PI3K pathway is extensively studied for its property in inhibiting apoptosis. PI3K is also known to regulate TLR-mediated inflammatory responses 22 . OxPAPC - TLR2 and TLR4 Inhibitor OxPAPC (1-palmitoyl-2-arachidonyl-sn-glycero-3-phosphorylcholine), is an oxidized phospholipid that has been shown to inhibit the signaling induced by bacterial lipopeptide and lipopolysaccharide (LPS). It acts by competing with CD14, LBP and MD2, the accessory proteins that interact with bacterial lipids, thus blocking the signaling of TLR2 and TLR4 23 . PD98059 - MAP Kinase Kinase Inhibitor PD98059 is a potent and selective inhibitor of MAP kinase kinase (also known as MAPK/ERK kinase or MEK kinase). It mediates its inhibitory properties by binding to the ERK-specific MAP kinase MEK, therefore preventing phosphorylation of ERK1/2 (p44/p42 MAPK) by MEK1/2. MAPK ERK1/2 is involved in TLR-induced production of cytokines 24 . 94 www.invivogen.com/inhibitors
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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
Page 44 and 45: ChemiComp E. coli ChemiComp E. coli
Page 46 and 47: 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 96 and 97: Pepinh-MYD - MyD88 inhibitory pepti
Page 98 and 99: 1. Martinon F,. et al., 2002. The i
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
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RNA INTERFERENCE 7 psiRNA-h7SK - 7S
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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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