Certificate of Analysis & Data Sheet
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 Human CD40L produced in E.Coli is a non-glycosylated, Polypeptide chain containing 149 amino acids and having a molecular mass of 16308 Dalton. Recombinant CD40L is purified by proprietary chromatographic techniques.
Sterile Filtered White lyophilized (freeze-dried) powder.
CD40L is lyophilized from a sterile concentrated solution (1mg/ml) with 10mM Sodium Phosphate pH=7.5.
It is recommended to reconstitute the lyophilized CD40L in sterile 18M-cm H2O not less than 100µg/ml, which can then be further diluted to other aqueous solutions containing carrier protein.
Lyophilized CD40L although stable at room temperature for 3 weeks, should be stored desiccated below -18 C. Upon reconstitution CD40L should be stored at 4 C between 2-7 days and for future use below -18 C. For long term storage it is recommended to add a carrier protein (0.1% HSA or BSA).
Please prevent freeze-thaw cycles.
Greater than 98.0% as determined by:
(a) Analysis by RP-HPLC.
(b) Anion-exchange FPLC.
(c) Analysis by reducing and non-reducing SDS-PAGE Silver Stained gel.
Amino Acid Sequence:
The sequence of the first five N-terminal amino acids was determined and was found to be Met-Gln-Lys-Gly-Asp.
Dimers and Aggregates:
Less than 1% as determined by silver-stained SDS-PAGE gel analysis.
PLR's CD40L is fully biologically active when compared to standard. The ED50 as determined by the dose-dependant stimulation of IL-12 & IL-8 induction by PMB (Peripheral Mononuclear) cells was found to be 5-10 ng/ml.
Less than 0.1 ng/µg (IEU/µg) of soluble CD40L.
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 soluble CD40L as a Reference Standard.
PLR's products are furnished for LABORATORY RESEARCH USE ONLY.