Human CD40 Ligand/TNFSF5 PE-conjugated Antibody

  • Species Reactivity
  • Specificity
    Detects human CD40 Ligand/TNFSF5 in direct ELISAs and Western blots. In direct ELISAs and Western blots, no cross-reactivity with recombinant human (rh) CD27 Ligand, rhCD30 Ligand, or recombinant mouse CD40 Ligand is observed.
  • Source
    Monoclonal Mouse IgG2B Clone # 40804
  • Purification
    Protein A or G purified from hybridoma culture supernatant
  • Immunogen
    E. coli-derived recombinant human CD40 Ligand/TNFSF5
    Accession # P29965
  • Formulation
    Supplied in a saline solution containing BSA and Sodium Azide.
  • Label
  • Flow Cytometry
    10 µL/106 cells
    See below
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 Human PBMCs by Flow Cytometry. Human peripheral blood mononuclear cells (PBMCs) treated with PMA and Calcium Ionomycin were stained with Mouse Anti-Human CD40 Ligand/TNFSF5 PE‑conjugated Monoclonal Antibody (Catalog # FAB617P, filled histogram) or isotype control antibody (Catalog # IC0041P, open histogram). View our protocol for Staining Membrane-associated Proteins.
Preparation and Storage
  • Shipping
    The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.
  • Stability & Storage
    Protect from light. Do not freeze.
    • 12 months from date of receipt, 2 to 8 °C as supplied.
Background: CD40 Ligand/TNFSF5
CD40 Ligand (CD40L), now renamed TNFSF5 but also known as CD154, TRAP and gp39, is a 34‑39 kDa type II transmembrane glycoprotein that belongs to the TNF superfamily (1‑3). Human CD40L is 261 amino acids (aa) in length and consists of a 22 aa cytoplasmic domain, a 24 aa transmembrane segment, and a 215 aa extracellular region that consists of multiple beta -strands and one N‑linked glycosylation site (4, 5). Although carbohydrates are present, they are not necessary for activity (6). As with other TNF superfamily members, CD40L will exist as a trimer, both as a membrane bound and soluble form (6‑8). The soluble form is 18 kDa in size and about 150 aa in length, and arises from intracellular proteolytic processing. As a trimer, the soluble form is bioactive (7‑9). Multiple mutations and alternate splice forms of CD40L exist, often associated with pathology and leading to truncated or nontrimerizable forms of CD40L (8). While CD40L is a ligand for CD40, the ligation of CD40L by CD40 initiates bidirectional signaling in both CD40 and CD40L expressing cells (10). The extracellular region of human CD40L is 99%, 88%, 86%, 82%, 75% and 75% aa identical to the extracellular regions of CD40L in rhesus monkey, bovine, porcine, canine, mouse and rat, respectively. CD40L binds to both CD40 and to integrin  alpha IIb beta 3 (CD41) (3, 11). In the cell membrane, it also associates with a unique splice variant of CD28 (CD28i) that may facilitate CD40L signal transduction (12). CD40L is expressed by monocytes, NK cells, mast cells, basophils, smooth muscle cells, endothelial cells, dendritic cells, activated and resting B cells, plus activated platelets and CD4+ T cells (13, 14). CD40L ligation of CD40 on dendritic cells (DC) initiates DC maturation and differentiation. CD40L signaling into naïve B cells promotes germinal center formation and isotope switching. With IL-21, CD40L generates IgA plus IgG3; with IL-4, CD40L generates IgG1 secretion (14, 15).
  • References:
    1. Zhang, G. (2004) Curr. Opin. Struct. Biol. 14:154.
    2. Hehlgans, T. and K. Pfeffer (2005) Immunology 115:1.
    3. Quezada, S.A. et al. (2004) Annu. Rev. Immunol. 22:307.
    4. Graf, D. et al. (1992) Eur. J. Immunol. 22:3191.
    5. Hollenbaugh, D. et al. (1992) EMBO J. 11:4313.
    6. Khandekar, S.S. et al. (2001) Prot. Exp. Purif. 23:301.
    7. Pietravalle, F. et al. (1996) J. Biol. Chem. 271:5965.
    8. Garber, E. et al. (1999) J. Biol. Chem. 274:33545.
    9. Vakkalanka, R.K. et al. (1999) Arthritis Rheum. 42:871.
    10. Eissner, G. et al. (2004) Cytokine Growth Factor Rev. 15:353.
    11. Prasad, K.S. et al. (2003) Proc. Natl. Acad. Sci. USA 100:12367.
    12. Mikolajczak, S.A. et al. (2004) J. Exp. Med. 199:1025.
    13. Lievens, D. et al. (2009) Thromb. Haemost. 102:206.
    14. Elgueta, R. et al. (2009) Immunol. Rev. 229:152.
    15. Avery, D.T. et al. (2008) J. Immunol. 181:1767.
  • 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
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Isotype Controls
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Mouse IgG2B PE-conjugated Antibody

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Staining Reagents
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Flow Cytometry Staining Buffer (1X)

Flow FC001 3
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Flow Cytometry Mouse Lyse Buffer (10X)

Flow FC003 1
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Flow Cytometry Human Lyse Buffer (10X)

Flow FC002 1
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