Recombinant Mouse soluble CD40 Ligand/TRAP

Synonyms CD40-L, Tumor necrosis factor ligand superfamily member 5, TNF-related activation protein, TRAP, T cell antigen Gp39, CD154 antigen, sCD40, IGM, IMD3, HIGM1, T-BAM, TNFSF5, hCD40L.

Introduction CD40L or CD154 is a membrane glycoprotein and differentiation antigen expressed on the surface of T-cells. The CD40 ligand stimulates B-cell proliferation and secretion of all immunoglobulin isotypes in the presence of cytokines . CD40 ligand has been shown to induce cytokine production and tumoricidal activity in peripheral blood monocytes. It also costimulates proliferation of activated T-cells and this is accompanied by the production of IFN-gamma , TNF-alpha , and IL2 .

Recombinant Mouse soluble CD40 Ligand/TRAPDescription sCD40 Mouse Recombinant produced in E.Coli is a non-glycosylated, Polypeptide chain containing 149 amino acids and having a molecular mass of 16409 Dalton.

The sCD40 is purified by proprietary chromatographic techniques.

Source Escherichia Coli.

Physical Appearance Sterile Filtered White lyophilized (freeze-dried) powder.

Recombinant Mouse soluble CD40 Ligand/TRAPFormulation Lyophilized from a sterile concentrated solution (1mg/ml) with 10mM Sodium Phosphate pH=7.5.

Solubility It is recommended to reconstitute the lyophilized sCD40 in sterile 18MΩ-cm H₂O not less than 100µg/ml, which can then be further diluted to other aqueous solutions.

Stability Lyophilized sCD40 although stable at room temperature for 3 weeks, should be stored desiccated below -18℃. Upon reconstitution CD154 should be stored at 4℃ between 2-7 days and for future use below -18℃.

For long term storage it is recommended to add a carrier protein (0.1% HSA or BSA).

Please prevent freeze-thaw cycles.

Purity Greater than 98.0% as determined by:

(a) Analysis by RP-HPLC.

(b) Analysis by SDS-PAGE.

Amino acid sequence The sequence of the first five N-terminal amino acids was determined and was found to be Met-Gln-Arg-Gly-Asp.

Biological Activity The ED50 as determined by its ability to induce MIP-1 alpha & TNF-alpha from mouse splenocytes was found to be 0.1 µg/ml corresponding to a Specific Activity of 10MIU/mg.

Recombinant Mouse soluble CD40 Ligand/TRAPProtein content Protein quantitation was carried out by two independent methods

1. UV spectroscopy at 280 nm using the absorbency value of 1.1 as the extinction coefficient for a 0.1% (1mg/ml) solution. This value is calculated by the PC GENE computer analysis program of protein sequences (IntelliGenetics).

2. Analysis by RP-HPLC, using a standard solution of sCD40 as a Reference Standard.