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.
Monoclonal Mouse IgG2B Clone # 40804
Protein A or G purified from hybridoma culture supernatant
E. coli-derived recombinant human CD40 Ligand/TNFSF5 Leu50-Leu261 Accession # P29965
Supplied in a saline solution containing BSA and Sodium Azide.
10 µL/106 cells
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.
Detection of CD40 Ligand/TNFSF5 in Human Peripheral Blood Monocytes by Flow Cytometry. Human peripheral blood monocytes stimulated with PMA and Ionomycin were stained with Mouse Anti-Human CD40 Ligand/TNFSF5 Fluorescein‑conjugated Monoclonal Antibody (Catalog # FAB617F, filled histogram) or isotype control antibody (Catalog # IC0041F, open histogram). View our protocol for Staining Membrane-associated Proteins.
Preparation and Storage
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 exists 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 alternative 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 shows 99%, 88%, 86%, 82%, 75% and 75% aa sequence identity 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 secretion; with IL-4, CD40L generates IgG1 secretion (14, 15).
Zhang, G. (2004) Curr. Opin. Struct. Biol. 14:154.
Hehlgans, T. and K. Pfeffer (2005) Immunology 115:1.
Quezada, S.A. et al. (2004) Annu. Rev. Immunol. 22:307.
Graf, D. et al. (1992) Eur. J. Immunol. 22:3191.
Hollenbaugh, D. et al. (1992) EMBO J. 11:4313.
Khandekar, S.S. et al. (2001) Protein Expr. Purif. 23:301.
Pietravalle, F. et al. (1996) J. Biol. Chem. 271:5965.
Garber, E. et al. (1999) J. Biol. Chem. 274:33545.
Vakkalanka, R.K. et al. (1999) Arthritis Rheum. 42:871.
Eissner, G. et al. (2004) Cytokine Growth Factor Rev. 15:353.
Prasad, K.S. et al. (2003) Proc. Natl. Acad. Sci. USA 100:12367.
Mikolajczak, S.A. et al. (2004) J. Exp. Med. 199:1025.
Lievens, D. et al. (2009) Thromb. Haemost. 102:206.
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.
The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.