Mouse CD40 Ligand/TNFSF5 Antibody

(5 citations)   
  • Species Reactivity
    Mouse
  • Specificity
    Detects mouse CD40 Ligand/TNFSF5 in Western blots.
  • Source
    Polyclonal Goat IgG
  • Purification
    Antigen Affinity-purified
  • 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
  • Western Blot
    0.1 µg/mL
    Recombinant Mouse CD40 Ligand/TNFSF5 (Catalog # 1163-CL)
  • Immunocytochemistry
    5-15 µg/mL
    See below
  • Neutralization
    Measured by its ability to neutralize CD40 Ligand/TNFSF5-induced proliferation in mouse splenic B cells. The Neutralization Dose (ND50) is typically 0.08-0.4 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
Immunocytochemistry
CD40 Ligand/TNFSF5 in Mouse Splenocytes. CD40 Ligand/TNFSF5 was detected in immersion fixed mouse splenocytes either stimulated with PHA (left panel) or untreated (right panel) using Goat Anti-Mouse CD40 Ligand/TNFSF5 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1163) at 15 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red; Catalog # NL001) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Non-adherent Cells.
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 Goat Anti-Mouse CD40 Ligand/TNFSF5 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1163). The ND50 is typically 0.08-0.4 µg/mL.
Preparation and Storage
  • Reconstitution
    Reconstitute at 0.2 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. Murine 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.

5 Citations: Showing 1 - 5
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Species
Applications
Sample Type
  1. Vaccination with a fusion protein that introduces HIV-1 gag antigen into a multitrimer CD40L construct results in enhanced CD8+ T cell responses and protection from viral challenge by vaccinia-gag.
    Authors: Gupta, Sachin, Termini, James M, Raffa, Francesc, Williams, Cindi-An, Kornbluth, Richard, Stone, Geoffrey
    J Virol, 2014;88(3):1492-501.
    Species: Mouse
    Sample Type: Cell Culture Supernates
    Application: WB
  2. The tumor necrosis factor family receptors RANK and CD40 cooperatively establish the thymic medullary microenvironment and self-tolerance.
    Authors: Akiyama T, Shimo Y, Yanai H, Qin J, Ohshima D, Maruyama Y, Asaumi Y, Kitazawa J, Takayanagi H, Penninger JM, Matsumoto M, Nitta T, Takahama Y, Inoue J
    Immunity, 2008;29(3):423-37.
    Species: Mouse
    Sample Type: Whole Tissue
    Application: IHC Paraffin-embedded
  3. CD40L expressed from the canarypox vector, ALVAC, can boost immunogenicity of HIV-1 canarypox vaccine in mice and enhance the in vitro expansion of viral specific CD8+ T cell memory responses from HIV-1-infected and HIV-1-uninfected individuals.
    Authors: Liu J, Yu Q, Stone GW, Yue FY, Ngai N, Jones RB, Kornbluth RS, Ostrowski MA
    Vaccine, 2008;26(32):4062-72.
    Species: Mouse
    Sample Type: Recombinant Protein
    Application: WB
  4. Preformed CD40 ligand exists in secretory lysosomes in effector and memory CD4+ T cells and is quickly expressed on the cell surface in an antigen-specific manner.
    Authors: Koguchi Y, Thauland TJ, Slifka MK, Parker DC
    Blood, 2007;110(7):2520-7.
    Species: Mouse
    Sample Type: Whole Cells
    Application: ICC
  5. Reduced pathology following infection with transgenic Leishmania major expressing murine CD40 ligand.
    Authors: Field AE, Wagage S, Conrad SM
    Infect. Immun., 2007;75(6):3140-9.
    Species: Human
    Sample Type: Cell Culture Supernates
    Application: ELISA Development
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