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IgG Antibodies InvivoGen offers a selection of monoclonal and polyclonal IgG antibodies. Most of them target pattern recognition receptors. These antibodies have been generated by immunization of mice or rats with DNA or recombinant proteins or peptides. These antibodies can be used for different applications. They have been tested in our laboratories for neutralization and/or flow cytometry, and for some of them for Western blot. MAb-TLR antibodies are also available conjugated with FITC. ANTIBODY DESCRIPTION REACTIVITY APPLICATIONS* QUANTITY CATALOG CODE Anti-Dectin-1 IgG MAb-mDectin-1 Mouse IgG2b (clone 2A11) Mouse Dectin-1 Neutralization, FC 100 μg mab-mdect Anti-Flagellin IgG Anti-Flagellin FliC Mouse IgG1 S. typhimurium flagellin WB 100 μg mabg-flic Anti-HA Tag IgG Anti-HA Tag Mouse IgG1 Influenza HA epitope WB, IP 250 μl ab-hatag Anti-TLR IgGs Anti-hTLR1-IgG Mouse IgG1 (H2G2) Human TLR1 Neutralization 100 μg mabg-htlr1 MAb-hTLR1 Mouse IgG1 (GD2.F4) Human TLR1 FC 100 μg mab-htlr1 MAb-hTLR1-FITC Mouse IgG1 (GD2.F4), FITC Human TLR1 FC 100 μg mab-htlr1f PAb hTLR1 Polyclonal rat IgG Human TLR1 Neutralization 200 μg pab-hstlr1 Anti-mTLR2-IgG Mouse IgG2a (C9A12) Mouse TLR2 Neutralization 100 μg mabg-mtlr2 MAb-hTLR2 Mouse IgG2a (TL2.1) Human TLR2 FC, IHC, WB 100 μg mab-htlr2 MAb-hTLR2-FITC Mouse IgG2a (TL2.1), FITC Human TLR2 FC, IHC 100 μg mab-htlr2f PAb hTLR2 Polyclonal rat IgG Human TLR2 Neutralization 200 μg pab-hstlr2 MAb-mTLR2 Mouse IgG1 (T2.5) Human/mouse TLR2 Neutralization , FC, IHC 100 μg mab-mtlr2 MAb-mTLR2-FITC Mouse IgG1 (T2.5), FITC Human/mouse TLR2 FC 100 μg mab-mtlr2f MAb-hTLR3 Mouse IgG1 (TLR3.7) Human TLR3 FC, WB 100 μg mab-htlr3 MAb-hTLR3-FITC Mouse IgG1 (TLR3.7), FITC Human TLR3 FC 100 μg mab-htlr3f MAb-hTLR4 Mouse IgG2a (HTA125) Human/monkey TLR4 FC, IHC 100 μg mab-htlr4 MAb-hTLR4-FITC Mouse IgG2a (HTA125), FITC Human/monkey TLR4 FC 100 μg mab-htlr4f PAb hTLR4 Polyclonal rat IgG Human TLR4 Neutralization 200 μg pab-hstlr4 MAb-mTLR4/MD2 Rat IgG2a (MTS510) Mouse TLR4/MD2 FC, IHC 100 μg mab-mtlr4md2 MAb-mTLR4/MD2-FITC Rat IgG2a (MTS510), FITC Mouse TLR4/MD2 FC 100 μg mab-mtlr4md2f PAb hTLR5 Polyclonal rat IgG Human TLR5 Neutralization 200 μg pab-hstlr5 Anti-mTLR5-IgG Rat IgG2a (Q23D11) Mouse TLR5 Neutralization 100 μg mabg-mtlr5 Anti-hTLR6-IgG Mouse IgG1 (C5C8) Human TLR6 Neutralization 100 μg mabg-htlr6 PAb hTLR6 Polyclonal rat IgG Human TLR6 Neutralization 200 μg pab-hstlr6 MAb-mTLR9 Mouse IgG2a (5G5) Human/mouse TLR9 FC, IHC, WB 100 μg mab-mtlr9 MAb-mTLR9-FITC Mouse IgG2a (5G5), FITC Human/mouse TLR9 FC 100 μg mab-mtlr9f * FC, flow cytometry; IHC, immunohistochemistry; IP, immunoprecipitation; WB, Western blot Contents and Storage All IgG antibodies are provided lyophilized. They are shipped at room temperature. Store at -20°C. www.invivogen.com/igg-antibody 125 ANTIBODIES & VACCINATION 5
ANTIBODIES & VACCINATION 5 Vaccine Adjuvants Adjuvants are essential for enhancing and directing the adaptative immune response to vaccine antigens. This response is mediated by two main types of lymphocytes, B and T cells. Upon activation by cytokines, B cells differentiate into memory B cells (long-lived antigen-specific B cells) or plasma cells (effector B cells that secrete large quantities of antibodies). Most antigens activate B cells using activated T helper (Th) cells, primarily Th1 and Th2 cells. Th1 cells secrete IFN-g, which activates macrophages and induces the production of opsonizing antibodies by B cells. The Th1 response leads mainly to a cell-mediated immunity (cellular response), which protects against intracellular pathogens (invasive bacteria, protozoa and viruses). The Th1 response activates cytotoxic T lymphocytes (CTL), a sub-group of T cells, which induce death of cells infected with viruses and other intracellular pathogens. Natural killer (NK) cells are also activated by the Th1 response, these cells play a major role in the induction of apoptosis in tumors and cells infected by viruses. Th2 cells secrete cytokines, including IL-4, which induces B cells to make neutralizing antibodies. Th2 cells generally induce a humoral (antibody) response critical in the defense against extracellular pathogens (helminthes, extracellular microbes and toxins). The magnitude and type of Th response to a vaccine can be greatly modulated through the use of adjuvants. For almost 80 years, aluminium salts (referred to as ‘alum’) have been the only adjuvant in use in human vaccines. Only in the last two decades, have novel adjuvants (MF59, AS04) been introduced in the formulation of new licensed vaccines. As our understanding of the mechanisms of ‘immunogenicity’ and ‘adjuvancy’ increases, new adjuvants and adjuvant formulations are being developed. Mechanisms of adjuvants Adjuvants may exert their effects through different mechanisms. Some adjuvants, such as alum and emulsions (e.g. MF59), function as delivery systems by generating depots that trap antigens at the injection site, providing slow release in order to continue the stimulation of the immune system. These adjuvants enhance the antigen persistence at the injection site and increase recruitment and activation of antigen presenting cells (APCs). Particulate adjuvants (e.g. alum) have the capability to bind antigens to form multi-molecular aggregates which will encourage uptake by APCs 1 . Some adjuvants are also capable of directing antigen presentation by the major histocompatibility complexes (MHC) 1 . Other adjuvants, essentially ligands for pattern recognition receptors (PRR), act by inducing the innate immunity, predominantly targeting the APCs and consequently influencing the adaptative immune response. Members of nearly all of the PRR families are potential targets for adjuvants. These include Toll-like receptors (TLRs), NOD-like receptors (NLRs), RIG-I-like receptors (RLRs) and C-type lectin receptors (CLRs). They signal through pathways that involve distinct adaptor molecules leading to the activation of different transcription factors. These transcription factors (NF-kB, IRF3) induce the production of cytokines and chemokines that play a key role in the priming, expansion and polarization of the immune responses. Activation of some members of the NLR family, such as NLRP3 and NLRC4, triggers the formation of a protein complex, called inflammasome, implicated in the induction of the pro-inflammatory cytokines IL-1b 2 and IL-18. The NLRP3 and NLRC4 inflammasomes have been involved in the innate immunity induced by certain adjuvants but their mechanism of action remains unclear. Alum & emulsions Alum is the most commonly used adjuvant in human vaccination. It is found in numerous vaccines, including diphtheria-tetanus-pertussis, human papillomavirus and hepatitis vaccines 3 . Alum provokes a strong Th2 response, but is rather ineffective against pathogens that require Th1–cellmediated immunity. Alum induces the immune response by a depot effect and activation of APCs. Recently, the NLRP3 inflammasome has been linked to the immunostimulatory properties of alum 2 although its role in adjuvantinduced antibody responses remains controversial. 126 www.invivogen.com/vaccine-adjuvants Emulsions (either oil-in-water or water-in-oil), such as Freund’s Incomplete Adjuvant (IFA) and MF59, can trigger depot generation and induction of MHC responses IFA induces a predominantly Th2 biased response with some Th1 cellular response. MF59 is a potent stimulator of both cellular (Th1) and humoral (Th2) immune responses 4 . However, the precise mode of action of emulsion-based adjuvants is still unclear. A complication with emulsion-based adjuvants is their potential to induce autoimmunity. PRR Ligands As classical adjuvants induce strong Th2 response with little or no Th1 response, the current challenge is to develop adjuvants which induce a strong Th1 bias important for vaccines against hepatitis, flu, malaria, and HIV. New adjuvants are being developed that are natural ligands or synthetic agonists for PRRs, either alone or with various formulations. PRR activation stimulates the production of pro-inflammatory cytokines/chemokines and type I IFNs that increase the host’s ability to eliminate the pathogen. Thus, the incorporation of pathogens associated molecular patterns (PAMPs) in vaccine formulations can improve and accelerate the induction of vaccinespecific responses. A number of these agonists are now in clinical or late preclinical stages of development for hepatitis and human papillomavirus vaccines 5, 6 . When used in combination with alum or classical emulsion adjuvants, the immune response can be biased towards a Th1 response 7 . TLR3 and RLR Ligands: Double-stranded RNA (dsRNA), which is produced during the replication of most viruses, is a potent inducer of innate immunity. Synthetic analogs of dsRNA, such as poly(I:C), have been tested as adjuvants. They act through TLR3 and RIG-I/MDA-5, inducing IL-12 and type I IFNs production, facilitating antigen cross-presentation to MHC class II molecules, and improving generation of cytotoxic T cells 8 . TLR4 Ligands: Bacterial lipopolysaccharides (LPS), which are ligands for TLR4, have long been recognized as potent adjuvants, but their pyrogenic activity have prevented their clinical use. The development of less toxic derivatives led to the production of MPLA (monophosphoryl lipid A), which formulated with alum (AS04) triggers a polarized Th1 response and is approved for clinical use in Europe 8 . TLR5 Ligands:The TLR5 ligand, bacterial flagellin, is a potent T-cell antigen and has potential as a vaccine adjuvant. Unlike other TLR agonists, flagellin tends to produce mixed Th1 and Th2 responses rather than strongly Th1 responses. Flagellin can be used as an adjuvant mixed with the antigen but it is more frequently fused to a recombinant vaccine antigen 9, 10 . TLR7/8 Ligands:The TLR7/8 pathway, specialized in the recognition of single stranded viral RNA, has demonstrated promising pre-clinical results as a target for potential vaccine adjuvants. Imidazoquinolines (i.e. imiquimod and R848) are synthetic componds that activate TLR7/8 in multiple subsets of dendritic cells leading to the production of IFN-a and IL-12 thus promoting a Th1 response 5 . TLR9 Ligands: Oligodeoxynucleotides containing specific CpG motifs (CpG ODNs) are recognized by TLR9. They enhance antibody production and strongly polarize Th cell responses to Th1 and away from Th2 responses 11,12 . NOD2 Ligands: Fragments of bacterial cell walls, such as muramyl dipeptide (MDP), have long been recognized as adjuvants. More recently, it was discovered that MDP triggers the activation of NOD2 and the NLRP3 inflammasome 13 . Adjuvants may be combined to achieve a stronger effect or a more potent skewing of immune responses. Currently, much effort is devoted to combining alum with TLR9 agoinsts 14 . In experimental models, administration of other combinations such as CpG ODNs with MDP or MPLA has proven very effective 15 . Adjuvants currently in clinical use enhance humoral responses but new adjuvants in clinical and preclinical trials are focused on generating multifaceted immune responses. The PRR pathways are an attractive source of novel adjuvants for vaccines due to their ability to induce strong cell-mediated immunity.
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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
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 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 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
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-
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