• Purity

  >95%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.

• Endotoxin Level

  <0.01 EU per 1 μg of the protein by the LAL method.  

• Activity

  Measured in a cell proliferation assay using enriched human B cells in the presence of IL-4. Spriggs, M.K.     et al. (1992) J. Exp. Med.     176:1543. The ED    ₅₀ for this effect is 0.2‑1.2 ng/mL in the presence of 20 ng/mL of Recombinant Human IL‑4 (Catalog # ) and a cross‑linking antibody, Mouse Anti-Hemagglutinin/HA Peptide Monoclonal Antibody (Catalog # ). Measured by its ability to induce IL-6 secretion by enriched human B cells. Urashima, M.     et al. (1996) Leukemia Res.     20:507. The ED    ₅₀ for this effect is approximately 0.5 ng/mL in the presence of Recombinant Human IL‑4 (Catalog # ) and a cross‑linking antibody, Mouse Anti‑Hemagglutinin/HA Peptide Monoclonal Antibody (Catalog # ).

• Source

  Human embryonic kidney cell, HEK293-derived

  Hemagglutinin tag YPYDVPDYA	GCN4-IZ (Met1-Ile29)	GGGSGGGSGGGS	Human CD40 Ligand (Met113-Leu261) Accession # P29965
  N-terminus			C-terminus

• Accession #

• N-terminal Sequence    
  Analysis

  Tyr

• Predicted Molecular Mass

  21.6 kDa

• SDS-PAGE

  22-26 kDa, reducing condtions

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 TNFSF, CD154, TRAP, and gp39, is a 34-39 kDa type II transmembrane glycoprotein that belongs to the TNF superfamily (1-3). Mature human CD40 Ligand consists of a 22 amino acid (aa) cytoplasmic domain, a transmembrane segment, and an 215 aa extracellular region (4, 5). The extracellular domain of human CD40 Ligand shares 74% and 76% aa sequence identity with mouse and rat CD40 Ligand, respectively. Similar to other TNF superfamily members, CD40 Ligand forms a bioactive homotrimer, both as membrane bound and soluble forms (6-9). The 18 kDa soluble form (aa 113-261) arises from proteolytic processing. Mutation and alternative splicing generate additional forms of CD40 Ligand that are often truncated or non-trimerizable (8). CD40 Ligand is expressed on platelets, as well as on activated T cells and B cells, basophils, eosinophils, fibroblasts, mast cells, monocytes, natural killer cells, vascular endothelial cells, and smooth muscle cells. CD40 Ligand binds to CD40, which is expressed on the surface of B cells, dendritic cells, macrophages, monocytes, platelets, endothelial, and epithelial cells (10). The interaction of CD40 Ligand with CD40 initiates signaling in both CD40 and CD40 Ligand expressing cells (11).  CD40 ligation by CD40 Ligand promotes B cell activation and T cell-dependent humoral responses (12, 13). CD40 Ligand dysregulation on T cells and antigen presenting cells contributes to the immune deficiency associated with HIV infection and AIDS (14, 15). It is also implicated in the pathology of multiple cardiovascular diseases including atherosclerosis, atherothrombosis, and restenosis (16, 17).

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

11. Eissner, G. et al. (2004) Cytokine Growth Factor. Rev. 15:353.

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

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

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

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

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

17. 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