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pZERO-TLR & pDeNy - Dominant Negative TLR & TLR Signaling Genes Features and Benefits DN TLR Genes Dominant negative (DN) TLR genes were generated by deleting a fragment of approximately 500 bp located at the 3’ end of each TLR gene corresponding to the TIR domain (TLR-DTIR genes). These truncated genes are still able to recognize their ligands but are unable to induce the signaling cascade. DN TLR genes are provided in the pZERO plasmid. pZERO-TLR plasmids are selectable with puromycin. HA-Tagged DN TLR Genes DN TLR genes are available with the influenza hemaglutinine (HA) tag fused at their 3’ end to facilitate their detection by Western blot. The HAtagged DN TLR proteins can be detected using the anti-HAtag antibody (see page 73). HA-tagged DN TLR genes are also provided in the pZERO plasmid. pZERO-TLR-HA plasmids are selectable with puromycin. DN TLR Signaling Genes DN forms of TLR signaling genes were created by inserting a mutation within the gene and/or deleting a region of the gene. These modifications have been described elsewhere (see table) and shown to block the wildtype form of these genes. DN TLR signaling genes are provided in the pDeNy plasmid which is selectable with Zeocin . pZERO or pDeNy plasmids can be cotransfected with a pUNO plasmid (see page 58). Selection of stable clones expressing both plasmids, pZERO/pUNO or pDeNy/pUNO, is achieved with addition of puromycin and blasticidin or Zeocin and blasticidin, respectively. Contents and Storage pZERO-TLR, pZERO-TLR-HA and pDeNy plasmids are provided as lyophilized transformed E. coli strains on paper disk. Transformed strains are shipped at room temperature and should be stored at -20ºC. Lyophilized E. coli cells are stable for at least 1 year when properly stored. pZERO-TLR and pZERO-TLR-HA plasmids are provided with 4 pouches of E. coli Fast-Media ® Puro, while pDeNy plasmids are provided with 4 pouches of E.coli Fast-Media ® Zeo (2 TB and 2 Agar). Related Products Puromycin, page 17 Zeocin , page 18 Anti-HAtag antibody, page 73 Fast-Media ® , page 45 PLASMID QTY www.invivogen.com/innate-immunity CAT. CODE (human) CAT. CODE (mouse) DN TLR Genes pZERO-TLR1 E. coli disk pzero-htlr1 pzero-mtlr1 pZERO-TLR2 E. coli disk pzero-htlr2 pzero-mtlr2 pZERO-TLR3 E. coli disk pzero-htlr3 pzero-mtlr3 pZERO-TLR4 E. coli disk pzero-htlr4 pzero-mtlr4 pZERO-TLR5 E. coli disk pzero-htlr5 pzero-mtlr5 pZERO-TLR6 E. coli disk pzero-htlr6 pzero-mtlr6 pZERO-TLR7 E. coli disk pzero-htlr7 pzero-mtlr7 pZERO-TLR8 E. coli disk pzero-htlr8 pzero-mtlr8 pZERO-TLR9 E. coli disk pzero-htlr9 pzero-mtlr9 pZERO-TLR10 E. coli disk pzero-htlr10 - HA-Tagged DN TLR Genes pZERO-TLR1-HA E. coli disk pzero-htlr1-ha pzero-mtlr1-ha pZERO-TLR2-HA E. coli disk pzero-htlr2-ha pzero-mtlr2-ha pZERO-TLR3-HA E. coli disk pzero-htlr3-ha pzero-mtlr3-ha pZERO-TLR4-HA E. coli disk pzero-htlr4-ha pzero-mtlr4-ha pZERO-TLR5-HA E. coli disk pzero-htlr5-ha pzero-mtlr5-ha pZERO-TLR6-HA E. coli disk pzero-htlr6-ha pzero-mtlr6-ha pZERO-TLR7-HA E. coli disk pzero-htlr7-ha pzero-mtlr7-ha pZERO-TLR8-HA E. coli disk pzero-htlr8-ha pzero-mtlr8-ha pZERO-TLR9-HA E. coli disk pzero-htlr9-ha pzero-mtlr9-ha pZERO-TLR10-HA E. coli disk pzero-htlr10-ha - PLASMID SPECIES MUTATION/ DELETION CAT. CODE DN TLR Signaling Genes pDeNy-hIRAK human aa1-211 pdn-hirak pDeNy-hMyD88 human aa161-296 pdn-hmyd88 pDeNy-mMyD88 mouse aa161-296 pdn-mmyd88 pDeNy-hNOD1 human aa127-518 pdn-hnod1 pDeNy-hNOD2 human L145P pdn-hnod2 pDeNy-hPKR human aa361-366 pdn-hpkr pDeNy-hRIG-I human aa1-217 pdn-hrigi pDeNy-hTIRAP human P125H pdn-htirap pDeNy-hTRAM human C117H pdn-htram pDeNy-hTRAF6 human aa289-522 pdn-traf6 pDeNy-hTRIF human aa387-566 + P434H pdn-htrif 63 INNATE IMMUNITY 3
Reporter Cell Lines Engineered HEK293 Cells Cells that constitutively express a given functional TLR gene are valuable tools for many applications, such as the study of the mechanisms involved in TLR recognition or signaling, and the development of new potential therapeutic drugs. InvivoGen provides HEK293 cells stably expressing human or murine TLR or NOD genes. • 293/TLR & 293/NOD Clones • HEK-Blue TLR and HEK-Blue NOD Cells Immune Reporter Cells InvivoGen provides THP1-XBlue , Ramos-Blue and RAW-Blue Cells, derived from a human monocytic cell line, a human B lymphocyte and murine macrophages, respectively. They naturally express a large repertoire of different classes of pattern recognition receptors (PRRs), such as the TLRs, NLRs and RLRs and stably express an NF-kB-inducible SEAP reporter gene to facilitate the study of these PRRs. • THP1-XBlue Cells • Ramos-Blue Cells • RAW-Blue Cells MEF Reporter Cell Lines InvivoGen provides murine embryonic fibroblasts (MEFs) isolated from various mouse strains and immortalized with the SV40 large antigen. They express a large repertoire of pattern recognition receptors. InvivoGen’s MEFs express an NF-kB-inducible reporter (SEAP) system allowing the convenient monitoring of NF-kB activation following TLR or RLR stimulation. • C3H/TLR4mut & C3H/WT MEFs • C57/WT MEFs Expression of TLR, NOD1/2 and RIG-I/MDA-5 mRNAs and their activity in InvivoGen’s Reporter Cell Lines* INNATE IMMUNITY 3+/- - +/- - +/- +/- - - - - +/- - - - 64 REPORTER CELL LINES TLR1 TLR2 TLR3 TLR4 TLR5 TLR6 TLR7 TLR8 TLR9 TLR10 NOD1 NOD2 RIG-I MDA-5 HEK293-Null cells THP1-XBlue Cells Ramos-Blue Cells RAW-Blue Cells C3H MEFs C57 MEFs + - + - + + - - - - + - - - + + +/- + + + + + + + + + + + + + - + + + - + - ND + ? ND ND + + + + + + + + + + + - + + +/- +/- + - - +/- + - + + + - ND ND + + + + +/- + + + + - +/- + + + + + +/- + - + + ND + - - + ND ND + + + + - + - - - - + + + + + + + + - + - - - - ND ND ND ND + + + + - + + + + - + + + + ND ND ND ND - ND ND ND ND - ND ND + + Rows with no background correspond to mRNA expression,rows with a grey background correspond to the activity B16-Blue IFN-a/b Cells InvivoGen provides B16-Blue IFN-a/b, a cell line that in addition to detect bioactive murine type I IFNs can also be used to study RIG-I/MDA-5 agonists, such as transfected poly(I:C) or 5’ppp-dsRNA (see page 106). www.invivogen.com/reporter-cells
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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
Page 14 and 15: SEAP Reporter Gene System Secreted
Page 16 and 17: HEK-Blue Detection - Real-Time Det
Page 18 and 19: Blasticidin S Description Blasticid
Page 20 and 21: LyoVec New formulation LyoVec is
Page 22 and 23: Poly-Arginine-HA Tagged Reprogrammi
Page 24 and 25: LENTI-Smart - Lentiviral Vector Pr
Page 26 and 27: Lentiviral Vector Production and Ce
Page 28 and 29: Antibody generation using pFUSE-CHI
Page 30 and 31: Engineering Fc regions for altered
Page 32 and 33: pBROAD & pWHERE - Optimized Vector
Page 34 and 35: pSELECT-Tag - Expression of a GFP-,
Page 36 and 37: pVIVO - Expression of Two Genes of
Page 38 and 39: pORF Description pORF plasmids feat
Page 40 and 41: Prom A-List - Human and Rodent Pro
Page 42 and 43: 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 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 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
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HEK-Blue IL-33/IL-1b Cells - IL-33
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HEK-Blue CD40L Cells - CD40L Repor
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5 ANTIBODIES & VACCINATION Antibodi
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Antibody generation using pFUSE-CHI
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Immunoglobulin A The mucosal surfac
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IgA Detection and Quantification In
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IgG Antibodies InvivoGen offers a s
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Mediators? 1. Leroux-Roels G., 2010
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Poly(I:C) Synthetic double-stranded
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pVAC - Induction of Neutralizing An
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CpG-Free DNA CpGs and the Immune Re
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pCpGfree-basic & pCpGfree-promoter
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pCpGfree-siRNA & pCpGfree-siRNADUO
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Custom-Made CpG-Free Genes InvivoGe
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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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