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.
PerCP (Peridinin-chlorophyll Protein Complex)
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 Activated Human Blood Lymphocytes by Flow Cytometry. Human peripheral blood lymphocytes treated with PMA and Calcium Ionomycin were stained with Mouse Anti-Human CD3 epsilon Fluorescein‑conjugated Monoclonal Antibody (Catalog # FAB100F) and either (A) Mouse Anti-Human CD40 Ligand/TNFSF5 PerCP‑conjugated Monoclonal Antibody (Catalog # FAB617C) or (B) Mouse IgG2B PerCP Isotype Control (Catalog # IC0041C). 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), also known as TNFSF5, CD154, TRAP and gp39, is a 33‑36 kDa type II transmembrane glycoprotein that belongs to the TNF superfamily. 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. Although carbohydrates are present, they are not necessary for activity. As with other TNF superfamily members, CD40L will exist as a trimer, both as a membrane bound and soluble form. The soluble form is 17-18 kDa in size and about 150 aa in length, and arises from intracellular proteolytic processing. As a trimer, the soluble form may be bioactive. Multiple mutations and alternate splice forms of CD40L exist, often associated with pathology and leading to truncated or nontrimerizable forms of CD40L. While CD40L is a ligand for CD40, the ligation of CD40L by CD40 initiates bidirectional signaling in both CD40 and CD40L expressing cells. 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 CD40 and to Integrin alpha IIb beta 3 (CD41), Integrin alpha 5 beta 1, and Mac-1. In the cell membrane, it also associates with a unique splice variant of CD28 (CD28i) that may facilitate CD40L signal transduction. 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. CD40L ligation of CD40 on dendritic cells (DC) initiates DC maturation and differentiation. CD40L signaling into naïve B cells promotes germinal center formation, plasma cell differentiation and isotope switching. With IL-21, CD40L generates IgA plus IgG3; with IL-4, CD40L generates IgE secretion.
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