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triggers NFAT activation in macrophages and dendritic cells. J Immunol. 178( 5): 3107-15. 10. Asai T. et al., 2003. Activation of transcription factors AP-1 and NF-κB in chronic cyclosporine A nephrotoxicity: role in beneficial effects of magnesium supplementation. Transplantation 75(7):1040–4. 11. Kang Y. et al., 2007. Calcineurin negatively regulates TLR-mediated activation pathways. J Immunol 179(7):4598–607. 12. Tigno-Aranjuez J. et al., 2010. Inhibition of RIP2’s tyrosine kinase activity limits NOD2-driven cytokine responses. Genes Dev. 24(23):2666-77. 13. Maemondo M et al. 2010. Gefitinib or Chemotherapy for Non–Small-Cell Lung Cancer with Mutated EGFR N Engl J Med. 362(25):2380-8. 14. Erridge C. et al., 2008. Oxidized Phospholipid Inhibition of Toll-like Receptor (TLR) Signaling Is Restricted to TLR2 and TLR4: roles for CD14, LPS-binding protein, and MD2 as targets for specificity of inhibition. J. Biol. Chem., 283: 24748 - 24759. 15. von Schlieffen E. et al., 2009. Multi-Hit Inhibition of Circulating and Cell-Associated Components of the Toll-Like Receptor 4 Pathway by Oxidized Phospholipids. Arterioscler Thromb Vasc Biol, 29: 356 - 362. 16. Loiarro M. et al., 2005. Peptide-mediated Interference of TIR Domain Dimerization in MyD88 Inhibits Interleukin-1-dependent Activation of NF-{kappa}B. J. Biol. Chem., 280: 15809-14. 17. Derossi D. et al., 1994. The third helix of the Antennapedia homeodomain translocates through biological membranes. J. Biol. Chem., 269: 10444-50. 18. Toshchakov VU. et al., 2005. Differential Involvement of BB Loops of Toll-IL-1 Resistance (TIR) Domain-Containing Adapter Proteins in TLR4- versus TLR2-Mediated Signal Transduction. J. Immunol., 175: 494 - 500. 19. Palmer JD, Rifkind D. 1974. Neutralization of the hemodynamic effects of endotoxin by polymyxin B. Surg Gynecol Obstet. 138(5):755-9. 20. Lindemann.RA 1988. Bacterial activation of human natural killer cells: role of cell surface lipopolysaccharide. Infect Immun. 56 (5): 1301–1308. 21. Duff GW, Atkins E. 1982. The inhibitory effect of polymyxin B on endotoxininduced endogenous pyrogen production. J Immunol Methods. 52(3):333-40. 22. Kuo CC. et al., 2006. Class I and III Phosphatidylinositol 3'-Kinase Play Distinct Roles in TLR Signaling Pathway. J. Immunol., 176: 5943 - 5949. 23 Lorne E. et al., 2009. Participation of Mammalian Target of Rapamycin Complex 1 in Toll-Like Receptor 2– and 4–Induced Neutrophil Activation and Acute Lung Injury. Am. J. Respir. Cell Mol. Biol., 41: 237 - 245. 24. Slee EA. et al., 1996. Benzyloxycarbonyl-Val-Ala-Asp (OMe) fluoromethylketone (Z-VAD.FMK) inhibits apoptosis by blocking the processing of CPP32. Biochem J. 315 ( Pt 1):21-4. 25. Dostert C. et al., 2009. Malarial hemozoin is a Nalp3 inflammasome activating danger signal. PLoS One. 4(8):e6510. Cytokine Receptor Inhibitor SB431542 - TGF-b Receptor Inhibitor SB431542 is a potent and selective inhibitor of transforming growth factor-b (TGF-b) superfamily type I activin receptor-like kinase (ALK) receptors, specifically ALK4, ALK5 and ALK7 1 . Inhibition of TGF-b signaling is known to induce the de-repression of epithelial fate and thus was hypothesized to benefit the reprogramming process. Indeed, treatment of OSKM-transduced human primary fibroblasts with a combination of SB431542 and PD0325901, a MEK inhibitor, was found to improve reprogramming efficiency of human cells 2 . 1. IInman GJ. et al., 2002. SB-431542 is a potent and specific inhibitor of transforming growth factor-beta superfamily type I activin receptor-like kinase (ALK) receptors ALK4, ALK5, and ALK7. Mol Pharmacol. 2002 Jul;62(1):65-74. 2. Lin T. et al., 2009. A chemical platform for improved induction of human iPSCs. Nat Methods. 6(11):805-8. Ion Channel Inhibitors Bafilomycin A1 - Proton pump inhibitor Bafilomycin A1 is a specific inhibitor of the lysosomal proton pump, thus it indirectly inhibits lysosomal enzymes which have acidic pH optima. Bafilomycin treatment has been shown to inhibit fusion of autophagosomes with both endosomes and lysosomes 1, 2 . Glybenclamide (glyburide) - Potassium channel inhibitor Glybenclamide, also known as glyburide, blocks the maturation of caspase-1 and pro-IL-1b by inhibiting the K+ efflux 3 . Glybenclamide was shown to potently block the activation of the NLRP3 inflammasome induced by PAMPs, DAMPs and crystalline substances 4 . Recent data suggest that glybenclamide works downstream of the P2X 7 receptor but upstream of NLRP3. 1. Yamamoto A. et al., 1998. Bafilomycin A1 prevents maturation of autophagic vacuoles by inhibiting fusion between autophagosomes and lysosomes in rat hepatoma cell line, H-4-II-E cells. Cell Struct Funct. 23(1):33-42. 2. Mousavi SA. et al., 2001. Effects of inhibitors of the vacuolar proton pump on hepatic heterophagy and autophagy. Biochim Biophys Acta. 1510(1-2):243-57 3. Laliberte RE. et al., 1999. ATP treatment of human monocytes promotes caspase-1 maturation and externalization. J Biol Chem. 274(52):36944-51. 4. Lamkanfi M. et al., 2009. Glyburide inhibits the Cryopyrin/Nalp3 inflammasome. J. Cell Biol., 187: 61 - 70. . Nuclear Export Inhibitor 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 cause the nuclear accumulation of proteins that shuttle between the cytosol and nucleus such as IRAK-1 and NLRC5 1, 2 . The cellular target of Leptomycin B has been identified as CRM1 (exportin 1), an evolutionarily conserved receptor for the nuclear export signal of proteins 3 . 1. Liu G. et al., 2008. Interleukin-1 receptor-associated kinase (IRAK)-1-mediated NF-kB activation requires cytosolic and nuclear activity. FASEB J 22(7): 2285-96. 2. Benko S. et al., 2010. NLRC5 limits the activation of inflammatory pathways. J Immunol. 185(3):1681-91. 3. Kudo N. et al., 1999. Leptomycin B inactivates CRM1/exportin 1 by covalent modification at a cysteine residue in the central conserved region. PNAS. 96(16):9112-7. NF-kB Activation Inhibitors 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 1 . TNF-a-dependent effects of NF-kB are important for TLR expression and cytokine production 2, 3 . 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 4, 5 . www.invivogen.com/inhibitors 155 INHIBITORS 8
INHIBITORS 8 Triptolide - NF-kB Inhibitor Triptolide, a diterpenoid isolated from the Chinese herb Tripterygium wilfordii hook F, has been used for centuries in traditional Chinese medicine to treat immune-related disorders. Current knowledge regarding its mechanism of action is still limited. So far, triptolide is known to block the activation of NF-kB 6, inhibit the heat shock response 7 and induce caspase activation 8 . 1. Pierce JW. et al., 1997. Novel Inhibitors of Cytokine-induced Ikappa Balpha Phosphorylation and Endothelial Cell Adhesion Molecule Expression Show Anti-inflammatory Effects in Vivo. J. Biol. Chem., 272: 21096. 2. Phulwani NK. et al., 2008. TLR2 Expression in Astrocytes Is Induced by TNF-{alpha}- and NF-{kappa}B-Dependent Pathways. J. Immunol., 181: 3841-3849. 3. Méndez-Samperio P. et al., 2008. Mycobacterium bovis Bacillus Calmette-Guérin Induces CCL5 Secretion via the Toll-Like Receptor 2-NF-{kappa}B and -Jun N-Terminal Kinase Signaling Pathways. Clin. Vaccine Immunol. 15: 277 - 283. 4 Sethi G. et al., 2007. Celastrol, a novel triterpene, potentiates TNF-induced apoptosis and suppresses invasion of tumor cells by inhibiting NF-kB–regulated gene products and TAK1-mediated NF-B activation. Blood, 109: 2727 - 2735. 5. Jung HW. et al., 2007. Celastrol inhibits production of nitric oxide and proinflammatory cytokines through MAPK signal transduction and NF-kappaB in LPS-stimulated BV-2 microglial cells. Exp Mol Med. 39(6):715-21. 6. Lee KY. et al., 1999. PG490 (triptolide) cooperates with tumor necrosis factor-alpha to induce apoptosis in tumor cells. J Biol Chem. 274: 13451-13455. 7. Westerheide SD. et al., 2006. Triptolide, an Inhibitor of the Human Heat Shock Response That Enhances Stress-induced Cell Death. J. Biol. Chem., 281: 9616 - 9622. 8. Carter BZ. et al., 2006. Triptolide induces caspase-dependent cell death mediated via the mitochondrial pathway in leukemic cells. Blood 108: 630 - 7. Mitogen-Activated Protein (MAP) Kinase Inhibitors 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) 1 . 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. PD0325901 - MEK Inhibitor PD0325901 is a selective inhibitor of mitogen-activated protein kinase kinase (MEK), with potential antineoplastic activity 2 . MEK is a key component of the RAS/RAF/MEK/ERK signaling pathway that is frequently activated in human tumors. This pathway plays also an important role in the self-renewing state of embryonic stem cells. Treatment with PD0325901 has been shown to enhance reprogramming efficiencies of induced pluripotent stem cells 3 . SB203580 - p38/RK MAP Kinase Inhibitor SB203580 is a pyridinyl imidazole inhibitor widely used to elucidate the roles of p38 mitogen-activated protein (MAP) kinase 4 . SB203580 inhibits also the phosphorylation and activation of protein kinase B (PKB, also known as Akt) 5 . SP600125 - JNK Inhibitor SP600125 is a potent, cell-permeable, selective and reversible inhibitor of c-Jun N-terminal kinase (JNK) 6 . It inhibits in a dose-dependent manner the phosphorylation of JNK. JNK is a member of the mitogen-activated protein kinase (MAPK) family and plays an essential role in TLR-mediated inflammatory responses. U0126 - MEK1 and MEK2 Inhibitor U0126 is a selective inhibitor of the MAP kinase kinases, MEK1 and MEK2. It acts by inhibiting the kinase activity of MEK1/2 thus preventing the activation of MAP kinases p42 and p44 which are encoded by the erk2 and erk1 genes respectively 7 . 1. Alessi DR. et al., 1995. PD 098059 is a specific inhibitor of the activation of mitogen-activated protein kinase kinase in vitro and in vivo. J. Biol. Chem. 270, 27489-27494. 2. Kohno M. & Pouyssegur J. 2006. Targeting the ERK signaling pathway in cancer therapy. Ann Med. 38(3):200-11. 3. Lin T. et al., 2009. A chemical platform for improved induction of human iPSCs. Nat Methods. 6(11):805-8. 4. Cuenda A. et al., 1995. SB 203580 is a specific inhibitor of a MAP kinase homologue which is stimulated by cellular stresses and interleukin-1. FEBS Lett. 364:229-233. 5. Lali FV. et al., 2000.The pyridinyl imidazole inhibitor SB203580 blocks phosphoinositide-dependent protein kinase activity, protein kinase B phosphorylation, and retinoblastoma hyperphosphorylation in interleukin-2-stimulated T cells independently of p38 mitogen-activated protein kinase. J Biol Chem. 275(10):7395-402. 6. Bennett BL. et al., 2001. SP600125, an anthrapyrazolone inhibitor of Jun N-terminal kinase. Proc. Natl. Acad. Sci. USA. 98:13681-13686. 7. Favata MF. et al., 1998. Identification of a novel inhibitor of mitogen-activated protein kinase.J. Biol. Chem. 273(29):18623-32. Phosphatidylinositol 3-Kinase (PI3K) Inhibitors 3-Methyladenine - PI3K inhibitor 3-Methyladenine is commonly used as a specific inhibitor of autophagic sequestration 1 . It blocks autophagy by inhibition of phosphatidylinositol 3-kinase (PI3K) activity, an enzyme required for autophagy 2 . LY294002 - PI3K Inhibitor LY294002 is a potent, cell permeable inhibitor of PI3K that acts on the ATP binding site of the enzyme 3 . LY294002 is often used to study the role of PI3K in apoptosis 4 and autophagy 2 . Wortmannin - PI3K Inhibitor Wortmannin is a cell-permeable, fungal metabolite that acts as a potent, selective and irrevesible inhibitor of PI3K 5 . Wortmannin has been used to determine the involvement of PI3K in many cellular processes, such as apoptosis 4 , autophagy 2 and TLR signaling 6 . 1. Seglen PO. & Gordon PB., 1982. 3-Methyladenine: specific inhibitor of autophagic/lysosomal protein degradation in isolated rat hepatocytes. PNAS. 1982 Mar;79(6):1889-92. 2. Blommaart EF. et al., 1997. The phosphatidylinositol 3-kinase inhibitors wortmannin and LY294002 inhibit autophagy in isolated rat hepatocytes. Eur. J. Biochem. 243: 240–246. 3. Vlahos CJ. et al., 1994. A specific inhibitor of phosphatidylinositol 3-kinase, 2-(4-morpholinyl)-8-phenyl-4H-1-benzopyran-4-one (LY294002). J. Biol. Chem 69(7):5241-8. 4. Duronio V., 2008. The life of a cell: apoptosis regulation by the PI3K/PKB pathway. Biochem J. 415(3):333-44. 5. Arcaro A. and Wymann MP., 1993. Wortmannin is a potent phosphatidylinositol 3-kinase inhibitor: the role of phosphatidylinositol 3,4,5-trisphosphate in neutrophil responses. Biochem. J. 296:297-301. 6. Fukao T and Koyasu S., 2003. PI3K and negative regulation of TLR signaling. Trends Immunol 24: 358-363. 156 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
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ChemiComp E. coli ChemiComp E. coli
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Fast-Media ® Preparation 1. Empty
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RIG-I-like Receptors 54 RLR & RLR-R
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Toll-Like Receptors Toll-Like Recep
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www.invivogen.com/innate-immunity 5
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IPAF and NAIP5 IPAF (NLRC4, CLAN/CA
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C-Type Lectin Receptors C-type lect
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pUNO pUNO-HA Toll-Like Receptors (T
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GENE NAME/ALIASES DESCRIPTION CAT.
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pDUO - Gene Associations Features a
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pZERO-TLR & pDeNy - Dominant Negati
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293/TLR & 293/NOD Clones 293/TLR Cl
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Response of HEK-Blue hTLR2 cells t
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Ramos-Blue Cells - NF-kB/AP-1 Repo
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MEF Cells C3H/TLR4mut & C3H/WT MEFs
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Antibodies for Detection ANTIBODY D
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- NF-kB binding site: Nuclear facto
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PRODUCT ORIGIN/DESCRIPTION TLR2 Ago
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PRODUCT ORIGIN/DESCRIPTION TLR9 Ago
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PRODUCT ORIGIN/DESCRIPTION Dectin-1
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LPS-PG (Porphyromonas gingivalis) R
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• FLA-ST ultrapure was purified b
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1. Zhao H. et al., 2004. Contributi
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1. Hornung V. et al., 2006. 5'-Trip
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TLR/NOD Ligand Screening InvivoGen
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Signal Transduction Inhibitors Invi
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Pepinh-MYD - MyD88 inhibitory pepti
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1. Martinon F,. et al., 2002. The i
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Nigericin - NLRP3 Inflammasome Indu
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Autophagy is a pathway by which cyt
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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
Page 149 and 150: RNA INTERFERENCE 7 Ready-Made psiRN
Page 151 and 152: RNA INTERFERENCE 7 Custom-Made psiR
Page 153 and 154: INHIBITORS 8 Inhibitors PRODUCT DES
Page 155: INHIBITORS 8 Pattern Recognition Re
Page 159 and 160: INHIBITORS 8 Heat Shock Protein Inh
Page 161 and 162: 9 CUSTOM SERVICES TLR/NOD Ligand Sc
Page 163 and 164: CUSTOM SERVICES 9 Custom Cloning :
Page 165 and 166: CUSTOM SERVICES 9 Prices 164 TERMS
Page 167 and 168: 166 PRODUCT (QUANTITY) CAT. CODE PA
Page 173 and 174: A 17-AAG 17-AAGH2 17-AEP-GA 2-Amino
Page 175 and 176: - ODN 4084-F - ODN INH-1 - ODN INH-
Page 177 and 178: ARGENTINA Genbiotech S.R.L. Tel: +5
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