• Purity

  >95%, by SDS-PAGE with silver staining

• Endotoxin Level

  <0.10 EU per 1 μg of the protein by the LAL method.  

• Activity

  Measured in a cell proliferation assay using mouse splenic B cells in the presence of IL-4. Banchereau, J.     et al. (1991) Science     251:70. The ED    ₅₀ for this effect is 0.07-0.35 ng/mL in the presence of a cross-linking antibody, Mouse Anti-Hemagglutinin/HA Peptide Monoclonal Antibody (Catalog # ).

• Source

  Chinese Hamster Ovary cell line, CHO-derived

  HA (YPYDVPDYA)	GCN4-IZ	GGGSGGGSGGGS	Mouse CD40 Ligand/TNFSF5  (Met112-Leu260) Accession # P27548
  N-terminus			C-terminus

• Accession #

• N-terminal Sequence    
  Analysis

  Tyr

• Predicted Molecular Mass

  22 kDa

• SDS-PAGE

  24-29 kDa, reducing conditions

Carrier Free

What does CF mean?

CF stands for Carrier Free (CF). We typically add Bovine Serum Albumin (BSA) as a carrier protein to our recombinant proteins. Adding a carrier protein enhances protein stability, increases shelf-life, and allows the recombinant protein to be stored at a more dilute concentration. The carrier free version does not contain BSA.

What formulation is right for me?

In general, we advise purchasing the recombinant protein with BSA for use in cell or tissue culture, or as an ELISA standard. In contrast, the carrier free protein is recommended for applications, in which the presence of BSA could interfere.

Background: CD40 Ligand/TNFSF5

CD40 Ligand, also known as TNFSF5, CD154, TRAP, or gp39, is a 33-39 kDa type II transmembrane glycoprotein member of the TNF superfamily (1, 2). Mature mouse CD40 Ligand consists of a 22 amino acid (aa) cytoplasmic domain, a transmembrane segment, and a 214 aa extracellular region (1). The extracellular domain of mouse CD40 Ligand shares 75% and 93% amino acid (aa) sequence identity with the human and rat proteins, respectively. CD40 Ligand is expressed as a homotrimer on platelets and activated T cells and B cells. It is up-regulated following stimulation of basophils, eosinophils, fibroblasts, mast cells, monocytes, natural killer cells, vascular endothelial cells, and smooth muscle cells. CD40 Ligand binds and activates CD40, which is expressed on the surface of B cells, dendritic cells, macrophages, monocytes, platelets, endothelial cells, and epithelial cells (3). The 18 kDa soluble form (aa 112-260)  arises from proteolytic processing and retains the ability to bind and activate CD40 (4, 5). Monomeric, dimeric, and trimeric forms of soluble CD40 Ligand bind to oligomeric CD40 on cell membranes (2). CD40 ligation by CD40 Ligand promotes B cell activation and T cell-dependent humoral responses (6, 7). CD40 Ligand dysregulation on T cells and antigen presenting cells contributes to the immune deficiency associated with HIV infection and AIDS (8, 9). It is also implicated in the pathology of multiple cardiovascular diseases including atherosclerosis, atherothrombosis, and restenosis (10, 11).

• References:

1. Armitage, R.J. et al. (1992) Nature 357:80.

2. Hollenbaugh, D. et al. (1992) EMBO J. 11:4313.

3. van Kooten, C. and J. Banchereau (1997) Curr. Opin. Immunol. 9:330.

4. Graf, D. et al. (1995) Eur. J. Immunol. 25:1749.

5. Mazzei, G.J. et al. (1995) J. Biol. Chem. 270:7025.

6. Rickert, R.C. et al. (2011) Immunol. Rev. 244:115.

7. Elgueta, R. et al. (2009) Immunol. Rev. 229:152.

8. Kornbluth, R.S. (2000) J. Leukoc. Biol. 68:373.

9. Chougnet, C. (2003) J. Leukoc. Biol. 74:702.

10. Pamukcu, B. et al. (2011) Ann. Med. 43:331.

11. Hassan, G.S. et al. (2012) Immunobiology 217:521.

• 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