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HEK-Blue CD40L Cells - CD40L Reporter Cells ➥ Study CD40L-CD40 interactions ➥ Screen for molecules that interfere with CD40L-CD40 cross-talk Background CD40 ligand (CD40L) is a member of the tumor necrosis factor (TNF) family of cell surface interaction molecules. It is mainly expressed in CD4+-T cells and interacts with CD40 on antigen-presenting cells to regulate both humoral and cellular immune responses. CD40L-CD40 interactions are thought to play an important role in the pathogenesis of certain diseases, including AIDS, Alzheimer’s disease and rheumatoid arthritis. Description HEK-Blue CD40L cells can serve to measure the bioactivity of CD40L through the secretion of embryonic alkaline phosphatase (SEAP) upon NF-kB activation. These cells were generated by stable transfection of HEK293 cells with the human CD40 gene and an NF-kB-inducible SEAP construct. The SEAP construct consists of the SEAP reporter gene under the control of the IFN-b minimal promoter fused to five NF-kB binding sites. Binding of CD40L to its receptor CD40 triggers a signaling cascade leading to the activation of NF-kB and the subsequent production of SEAP (figure 1). CD40L-CD40 interaction can be monitored by assessing the levels of SEAP using a SEAP detection assay, such as QUANTI-Blue (figure 2). HEK293 cells express endogenously the receptors for the cytokines IL-1b and TNF-a which share a common signaling pathway with CD40L. Consequently, HEK-Blue CD40L cells also respond to IL-1b and TNF-a. IL-1b- and TNF-a-mediated SEAP production can be blocked using neutralizing antibodies, such as anti-hIL-1b-IgA and anti-hTNF-a-IgA antibodies respectively (see page 104). • Detection range for CD40L: 5 ng - 1 μg/ml HEK-Blue CD40L cells are resistant to the selective antibiotics blasticidin and Zeocin . Contents and Storage HEK-Blue CD40L cells are grown in DMEM medium, 2mM L-glutamine, 10% FBS supplemented with 100 μg/ml Zeocin and 30 μg/ml blasticidin. Each vial contains 5-7 x 10 6 cells and is supplied with 10 mg Zeocin , 1 mg blasticidin, 50 mg Normocin and 1 pouch of QUANTI-Blue . Cells are shipped on dry ice. HEK-Blue CD40L cells are also available in a kit, see below: Figure 2: Stimulation of HEK-Blue CD40L cells by human CD40L was assessed by measuring the levels of SEAP using QUANTI-Blue . HEK-Blue CD40L Kit • HEK-Blue CD40L Cells 5-7 x 10 6 cells • Anti-hCD40L-IgA antibody 100 μg • QUANTI-Blue 1 pouch (100 ml) • Normocin 1 ml (50 mg/ml) • Blasticidin 100 μl (10 mg/ml) • Zeocin 100 μl (100 mg/ml) Figure 1: CD40L-induced NF-kB signaling pathway. PRODUCT QUANTITY CAT. CODE HEK-Blue CD40L Cells 5-7 x 10 6 cells hkb-cd40 HEK-Blue CD40L Kit 1 kit hkb-cd40-kit www.invivogen.com/cytokine-sensor-cells 115 CYTOKINE SIGNALING 4
CYTOKINE SIGNALING 4 HEK-Blue Cytokine Kits PRODUCT CONTENTS CAT. CODE HEK-Blue IL-4/IL-13 Kit • HEK-Blue IL-4/IL-13 Cells (5-7 x 10 6 cells) • Normocin (1 ml @ 50 mg/ml) • Anti-hIL-4-IgA neutralizing antibody (100 μg) • Blasticidin (100 μl @ 10 mg/ml) • Anti-hIL-13-IgA neutralizing antibody (100 μg) • Zeocin (100 μl @ 100 mg/ml) • QUANTI-Blue (1 pouch, 100 ml) HEK-Blue IL-6 Kit • HEK-Blue IL-6 Cells (5-7 x 10 6 cells) • Normocin (1 ml @ 50 mg/ml) • Anti-hIL-6-IgA neutralizing antibody (100 μg) • HygroGold (100 μl @ 100 mg/ml) • QUANTI-Blue (1 pouch, 100 ml) • Zeocin (100 μl @ 100 mg/ml) HEK-Blue TNF-a/IL-1b Kit • HEK-Blue TNF-a/IL-1b Cells (5-7 x 10 6 cells) • QUANTI-Blue (1 pouch, 100 ml) • Anti-hTNF-a-IgA neutralizing antibody (100 μg) • Normocin (1 ml @ 50 mg/ml) • Anti-hIL-1b-IgA neutralizing antibody (100 μg) • Zeocin (100 μl @ 100 mg/ml) HEK-Blue IL-1b Kit • HEK-Blue IL-1b Cells (5-7 x 10 6 cells) • Normocin (1 ml @ 50 mg/ml) • Anti-hIL-1b-IgA neutralizing antibody (100 μg) • HygroGold (100 μl @ 100 mg/ml) • QUANTI-Blue (1 pouch, 100 ml) • Zeocin (100 μl @ 100 mg/ml) HEK-Blue IL-18/IL-1b Kit • HEK-Blue IL-18/IL-1b Cells (5-7 x 10 6 cells) • Normocin (1 ml @ 50 mg/ml) • Anti-hIL-18-IgA neutralizing antibody (100 μg) • Blasticidin (100 μl @ 10 mg/ml) • Anti-hIL-1b-IgA neutralizing antibody (100 μg) • HygroGold (100 μl @ 100 mg/ml) • QUANTI-Blue (1 pouch, 100 ml) • Zeocin (100 μl @ 100 mg/ml) HEK-Blue TGF-b Kit • HEK-Blue TGF-b Cells (5-7 x 10 6 cells) • Blasticidin (100 μl @ 10 mg/ml) • Anti-hTGF-b-IgA neutralizing antibody (100 μg) • HygroGold (100 μl @ 100 mg/ml) • QUANTI-Blue (1 pouch, 100 ml) • Zeocin (100 μl @ 100 mg/ml) • Normocin (1 ml @ 50 mg/ml) HEK-Blue CD40L Kit • HEK-Blue CD40L Cells (5-7 x 10 6 cells) • Normocin (1 ml @ 50 mg/ml) • Anti-hCD40L-IgA neutralizing antibody (100 μg) • Blasticidin (100 μl @ 10 mg/ml) • QUANTI-Blue (1 pouch, 100 ml) • Zeocin (100 μl @ 100 mg/ml) Contents and Storage 116 www.invivogen.com/cytokine-sensor-cells hkb-stat6-kit hkb-il6-kit hkb-tnfil1-kit hkb-il1b-kit hkb-il18-kit hkb-tgfb-kit hkb-cd40-kit HEK-Blue Cytokine cells are shipped on dry ice. Thaw immediately or store in liquid nitrogen. Other products are shipped at room temperature and should be stored at -20°C.
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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
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
Page 114 and 115: HEK-Blue IL-33/IL-1b Cells - IL-33
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 and 156: INHIBITORS 8 Pattern Recognition Re
Page 157 and 158: INHIBITORS 8 Triptolide - NF-kB Inh
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
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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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