Human CD40/TNFRSF5 Antibody

(6 citations)   
  • Species Reactivity
    Human
  • Specificity
    Detects human CD40 in direct ELISAs and Western blots. In direct ELISAs and Western blots, no cross-reactivity with recombinant human (rh) 4‑1BB, rhBAFF R, rhCD27, rhCD30, rhDR3, rhDR6, rhEDAR, rhFas, rhGITR, rhHVEM, rhNGF R, rhOPG, rhRANK, rhTAJ, rhTNF RI, or rhTNF RII is observed.
  • Source
    Monoclonal Mouse IgG2B Clone # 82102
  • Purification
    Protein A or G purified from ascites
  • Immunogen
    Mouse myeloma cell line NS0-derived recombinant human CD40/TNFRSF5
    Glu21-Arg193
    Accession # P25942
  • Formulation
    Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied as a 0.2 µm filtered solution in PBS.
  • Endotoxin Level
    <0.10 EU per 1 μg of the antibody by the LAL method.
  • Label
    Unconjugated
Applications
  •  
    Recommended
    Concentration
    Sample
  • Western Blot
    2 µg/mL
    See below
  • Neutralization

    Measured by its ability to neutralize CD40 Ligand/TNFSF5-induced proliferation in human B cell enriched peripheral blood mononuclear cells (PBMC). At 5 µg/mL, this anti-hCD40 antibody will neutralize approximately 80% of 10 µg/mL Recombinant Human CD40 Ligand/TNFSF5 -induced proliferation of human B-cell enriched PBMCs.

     

     

Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.
Data Examples
Cell Proliferation Induced by CD40 Ligand/TNFSF5 and Neutralization by Human CD40/TNFRSF5 Antibody. In the presence of Recombinant Human IL‑4 (Catalog # 204-IL), Recombinant Human CD40 Ligand/TNFSF5 (Catalog # 6245-CL) stimulates proliferation in human B cell enriched peripheral blood mononuclear cells (PBMC) in a dose-dependent manner (orange line). Proliferation elicited by Recombinant Human CD40 Ligand/TNFSF5 (10 µg/mL) is neutralized (green line) by increasing concentrations of Human CD40/TNFRSF5 Monoclonal Antibody (Catalog # MAB6322). At 5 µg/mL, this anti-hCD40 antibody will neutralize approximately 80% of Recombinant Human CD40 Ligand/TNFSF5 -induced proliferation in the presence of Recombinant Human IL‑4 (20 ng/mL).
Detection of CD40/TNFRSF5 Mouse by Western Blot. Western blot shows lysates of human spleen tissue. PVDF membrane was probed with 2 µg/mL of Mouse Anti-Human CD40/TNFRSF5 Monoclonal Antibody (Catalog # MAB6322) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF007). A specific band was detected for CD40/TNFRSF5 at approximately 40 kDa (as indicated). This experiment was conducted under non-reducing conditions and using Immunoblot Buffer Group 1.
Preparation and Storage
  • Reconstitution
    Reconstitute at 0.5 mg/mL in sterile PBS.
  • Shipping
    The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
  • Stability & Storage
    Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
    • 12 months from date of receipt, -20 to -70 °C as supplied.
    • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
    • 6 months, -20 to -70 °C under sterile conditions after reconstitution.
Background: CD40/TNFRSF5

CD40 is a type I transmembrane glycoprotein belonging to the TNF receptor superfamily. Mature human CD40 consists of a 172 amino acid (aa) extracellular domain, a 22 aa transmembrane region and a 62 aa cytoplasmic domain (1). Human and mouse CD40 share 62% aa identity. CD40 is expressed in B cells, follicular dendritic cells, dendritic cells, activated monocytes, macrophages, endothelial cells, vascular smooth muscle cells, and several tumor cell lines (2). The extracellular domain has the cysteine-rich repeat regions, which are characteristic for many of the receptors of the TNF superfamily. Interaction of CD40 with its ligand, CD40L, leads to aggregation of CD40 molecules, which in turn interact with cytoplasmic components to initiate signaling pathways. Early studies on the CD40-CD40L system revealed its role in humoral immunity. Interaction between CD40L on T cells and CD40 on B cells stimulated B cell proliferation and provided the signal for immunoglobulin isotype switching (3). Mutations in the CD40L gene, which resulted in a CD40L molecule unable to interact with CD40, are responsible for the hyper-IgM syndrome (4). Cross-linking of CD40 with antibodies or by CD40 binding to CD40L produces cell type-specific responses which include costimulation and induction of proliferation, induction of cytokine production, rescue from apoptosis, and upregulation of adhesion molecules (5). Some of the early events of intracellular signaling by the CD40-CD40L system include the association of the CD40 with TRAFs and the activation of various kinases (6-8).

  • References:
    1. Torres, R.M. and E.A. Clark (1992) J. Immunol. 148:620.
    2. Schonbeck, U. et al. (1997) J. Biol. Chem. 272:19569.
    3. Armitage, R.J. et al. (1993) J. Immunol. 150:3671.
    4. Callard, R.E. et al. (1993) Immunol. Today 14:559.
    5. Stout, R.D. and J. Suttles (1996) Immunol. Today 17:487.
    6. Pullen, S.S. et al. (1999) Biochemistry 38:10168.
    7. Faris, M. et al. (1994) J. Exp. Med. 179:1923.
    8. Hanissian, S.H. and R.S Geha (1997) Immunity 6:379.
  • Entrez Gene IDs:
    958 (Human); 21939 (Mouse); 171369 (Rat); 102118696 (Cynomolgus Monkey)
  • Alternate Names:
    B cell surface antigen CD40; B-cell surface antigen CD40; Bp50B cell-associated molecule; CD40 antigen; CD40 molecule, TNF receptor superfamily member 5; CD40 type II isoform; CD40; CD40L receptor; CDw40; MGC9013; nerve growth factor receptor-related B-lymphocyte activation molecule; p50; TNFRSF5; TNFRSF5CD40 antigen (TNF receptor superfamily member 5); tumor necrosis factor receptor superfamily member 5; tumor necrosis factor receptor superfamily, member 5
Related Research Areas
Citations:

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

6 Citations: Showing 1 - 6
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Species
Applications
Sample Type
  1. Circulating CXCR5(+)CXCR3(+)PD-1(lo) Tfh-like cells in HIV-1 controllers with neutralizing antibody breadth
    Authors: E Martin-Gay, J Cronin, T Hickman, Z Ouyang, M Lindqvist, KE Kolb, J Schulze Zu, R Cubas, F Porichis, AK Shalek, J van Lunzen, EK Haddad, BD Walker, DE Kaufmann, M Lichterfel, XG Yu
    JCI Insight, 2017;2(2):e89574.
    Species: Human
    Sample Type: Whole Cells
    Application: Neut
  2. Up-Regulated Expression of Matrix Metalloproteinases in Endothelial Cells Mediates Platelet Microvesicle-Induced Angiogenesis
    Authors: C Sun, SB Feng, ZW Cao, JJ Bei, Q Chen, WB Zhao, XJ Xu, Z Zhou, ZP Yu, HY Hu
    Cell. Physiol. Biochem., 2017;41(6):2319-2332.
    Species: Human
    Sample Type: Whole Cells
    Application: Neut
  3. A soluble form of IL-27Ralpha is a natural IL-27 antagonist.
    Authors: Dietrich C, Candon S, Ruemmele F, Devergne O
    J Immunol, 2014;192(11):5382-9.
    Species: Human
    Sample Type: Whole Cells
    Application: Neut
  4. ICOS-LICOS interaction is critically involved in TGN1412-mediated T-cell activation.
    Blood, 2012;119(26):6268-77.
    Species: Human
    Sample Type: Whole Cells
    Application: Neut
  5. Circulating myeloid dendritic cells of advanced cancer patients result in reduced activation and a biased cytokine profile in invariant NKT cells.
    Authors: van der Vliet HJ, Wang R, Yue SC, Koon HB, Balk SP, Exley MA
    J. Immunol., 2008;180(11):7287-93.
    Species: Human
    Sample Type: Whole Cells
    Application: Neut
  6. IL-21 and BAFF/BLyS synergize in stimulating plasma cell differentiation from a unique population of human splenic memory B cells.
    Authors: Ettinger R, Sims GP, Robbins R, Withers D, Fischer RT, Grammer AC, Kuchen S, Lipsky PE
    J. Immunol., 2007;178(5):2872-82.
    Species: Human
    Sample Type: Whole Cells
    Application: Neut
Isotype Controls
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Mouse IgG2B Isotype Control

Ctrl MAB004 32  
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Mouse IgG2B Isotype Control

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Mouse IgG2B Isotype Control

Ctrl MAB0042 6
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Secondary Antibodies
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Mouse IgG HRP-conjugated Antibody

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Mouse F(ab)2 IgG (H+L) PE-conjugated Antibody

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Donkey Anti-Mouse IgG NorthernLights™ NL557-conjugate Antibody

Flow, IHC, ICC NL007 9
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Donkey Anti-Mouse IgG Biotinylated Antibody

WB BAF018 1
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Mouse IgG HRP-conjugated Antibody

WB HAF018 9  
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Donkey Anti-Mouse IgG NorthernLights™ NL637-conjugated Antibody

Flow, IHC, ICC NL008 8
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Mouse F(ab)2 IgG (H+L) APC-conjugated Antibody

Flow F0101B 2  
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Donkey Anti-Mouse IgG NorthernLights™ NL493-conjugated Antibody

Flow, IHC, ICC NL009 5
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Goat Anti-Mouse IgG Biotinylated Antibody

WB BAF007 4
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Mouse IgG Antibody

WB AF007 4
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Mouse F(ab)2 IgG (H+L) Fluorescein-conjugated Antibody

Flow F0103B 5  
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Mouse/Rabbit IgG VisUCyte HRP Polymer Antibody

IHC VC002  
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Mouse IgG VisUCyte HRP Polymer Antibody

IHC VC001  
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Rat Anti-Mouse IgG2B APC-conjugated Antibody

Flow F0133 1  
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Goat Anti-Mouse IgG Fc Affinity Purified Polyclonal Ab

G-202-C 2
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Mouse F(ab)2 IgG (H+L) PerCP-conjugated Antibody

Flow F0114  
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Mouse IgG2B Antibody

Flow MAB0043  
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Rat Anti-Mouse IgG2B PE-conjugated Antibody

Flow F0132  
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Rat Anti-Mouse IgG2B Fluorescein-conjugated Antibody

Flow F0134 1
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