Mouse CD40 Ligand/TNFSF5 Antibody

(1 citations)   
  • Species Reactivity
    Mouse
  • Specificity
    Detects mouse CD40 Ligand in direct ELISAs. In direct ELISAs, no cross-reactivity with recombinant mouse (rm) CD27 Ligand, rmCD30 Ligand, or recombinant human CD40 Ligand is observed.
  • Source
    Monoclonal Rat IgG2A Clone # 208109
  • Purification
    Protein A or G purified from hybridoma culture supernatant
  • Immunogen
    Mouse myeloma cell line NS0-derived recombinant mouse CD40 Ligand/TNFSF5
    Glu61-Leu260
    Accession # P27548
  • 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
  • Flow Cytometry
    0.25 µg/106 cells
    See below
  • CyTOF-ready
    Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.
  • Neutralization
    Measured by its ability to neutralize CD40 Ligand/TNFSF5-induced proliferation in mouse splenic B cells. The Neutralization Dose (ND50) is typically 0.0075-0.045 µg/mL in the presence of 0.5 ng/mL Recombinant Mouse CD40 Ligand/TNFSF5.
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
Detection of CD40 Ligand/TNFSF5 in Mouse Splenocytes by Flow Cytometry. Mouse splenocyteseither (A) stimulated to induce Th1 cells or (B) unstimulated were stained with Rat Anti-Mouse CD40 Ligand/TNFSF5 Monoclonal Antibody (Catalog # MAB1163) followed by Allophycocyanin-conjugated Anti-Rat IgG Secondary Antibody (Catalog # F0113) and Rat Anti-Mouse CD4 PE‑conjugated Monoclonal Antibody (Catalog # FAB554P). Quadrant markers were set based on control antibody staining (Catalog # MAB006). View our protocol for Staining Membrane-associated Proteins.
Proliferation Induced by CD40 Ligand/TNFSF5 and Neutralization by Mouse CD40 Ligand/TNFSF5 Antibody. Recombinant Mouse CD40 Ligand/TNFSF5 (Catalog # 8230-CL) induces proliferation in mouse splenic B cells in a dose-dependent manner (orange line), as measured by Resazurin (Catalog # AR002). Proliferation elicited by Recombinant Mouse CD40 Ligand/TNFSF5 (0.5 ng/mL) is neutralized (green line) by increasing concentrations of Rat Anti-Mouse CD40 Ligand/TNFSF5 Monoclonal Antibody (Catalog # MAB1163). The ND50 is typically
0.0075-0.045 µg/mL.
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 Ligand/TNFSF5

CD40 ligand (CD40L; also known as CD154, TNFSF5, TRAP, or gp39) is a 260 amino acid (aa) type II transmembrane glycoprotein belonging to the TNF family. Mouse CD40L consists of a 22 aa cytoplasmic domain, a 24 aa transmembrane domain, and 214 aa extracellular domain bearing a single glycosylation site (1, 2). CD40L is expressed predominantly on activated CD4+ T lymphocytes and also found in other types of cells, including NK cells, mast cells, basophils, and eosinophils. Murine CD40L shares 78% amino acid sequence identity with human CD40L. Native bioactive soluble CD40L exists. Soluble human trimeric CD40L secreted by stimulated T cells has been shown to be generated by proteolysis in the microsomes (3). Both membrane bound and soluble CD40L induce similar effects on B cells (3, 4). The receptor of CD40L is CD40, a type I transmembrane glycoprotein belonging to the TNF receptor family. CD40 is expressed on B lymphocytes, monocytes, dendritic cells, and thymic epithelium. Although all monomeric, dimeric, and trimeric forms of soluble CD40L can bind to CD40, the soluble trimeric form of CD40L has the most potent biological activity through oligomerization of cell surface CD40, a common feature of TNF receptor family members (2). The genetic defect in the hyper-IgM syndrome is due to point mutations or deletions of the gene encoding the CD40L, which prevent CD40L from interacting with CD40 (5‑7). CD40L mediates a range of activities on B cells including induction of activation-associated surface antigen, entry into the cell cycle, isotype switching, Ig secretion, and memory generation (8, 9). CD40-CD40L interaction also plays important roles in monocyte activation and dendritic cell maturation (10).

  • References:
    1. Armitage, R.J. et al. (1992) Nature 357:80.
    2. Hollenbaugh, D. et al. (1992) EMBO J. 11:4313.
    3. Fabienne, P. et al. (1996) J. Biol. Chem. 271:5965.
    4. Fabienne, P. et al. (1996) Eur. J. Immunol 26:725.
    5. Arrufo, A. et al. (1993) Cell 72:291.
    6. Hill, A. and N. Chapel et al. (1993) Nature 361:494.
    7. Korthauer, U. et al. (1993) Nature 361:539.
    8. Spriggs, M.K. et al. (1992) J. Exp. Med. 176:1543.
    9. Fanslow, W.C. et al. (1994) Seminars in Immunology 6:267.
    10. Kooten, C.V. and J. Banchereau (2000) J. Leukoc. Biol. 67:2.
  • Entrez Gene IDs:
    959 (Human); 21947 (Mouse); 84349 (Rat)
  • Alternate Names:
    CD154 antigen; CD154; CD40 antigen ligand; CD40 Ligand; CD40-L; CD40LG; CD40LIGM; gp39; hCD40L; HIGM1; T-B cell-activating molecule; T-BAM; T-cell antigen Gp39; TNF-related activation protein; TNFSF5; TNFSF5IMD3; TRAP; TRAPtumor necrosis factor (ligand) superfamily, member 5 (hyper-IgM syndrome); tumor necrosis factor (ligand) superfamily member 5; Tumor necrosis factor ligand 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.

1 Citations: Showing 1 - 1

  1. Role of Valpha14+ NKT cells in the development of Hepatitis B virus-specific CTL: activation of Valpha14+ NKT cells promotes the breakage of CTL tolerance.
    Authors: Ito H, Ando K, Ishikawa T
    Int. Immunol., 2008;20(7):869-79.
    Species: Mouse
    Sample Type: In Vivo
    Application: In vivo
Isotype Controls
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Rat IgG2A Isotype Control

Ctrl MAB006 90  
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Normal Rat IgG Control

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Normal Rat IgG Control (Azide Free)

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Rat IgG2A Isotype Control

Ctrl MAB006R  
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Secondary Antibodies
Description Application Cat# Citations Images  

Rat IgG HRP-conjugated Antibody

WB, Simple Western HAF005 7  
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Goat Anti-Rat F(ab)2 IgG (H+L)
APC-conjugated Antibody

Flow F0113 2  
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Goat Anti-Rat IgG NorthernLights™ NL557-conjugated Antibody

Flow, IHC, ICC NL013 2
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Rat F(ab)2 IgG (H+L) PE-conjugated Antibody

Flow F0105B 2  
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Goat Anti-Rat IgG NorthernLights™ NL493-conjugated Antibody

Flow, IHC, ICC NL015 1
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Goat Anti-Rat IgG Biotinylated Antibody

WB BAF005 2
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Goat Anti-Rat IgG Antibody

WB AF005 1
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Rat IgG VisUCyte HRP Polymer Antibody

IHC VC005  
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Rat F(ab)2 IgG (H+L) Fluorescein-conjugated Antibody

Flow F0104B  
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Goat Anti-Rat IgG NorthernLights™ NL637-conjugated Antibody

Flow, IHC, ICC NL014
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Goat Anti-Rat F(ab)2 IgG (H+L) PerCP-conjugated Antibody

Flow F0115 1
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