Optimized DNA sequence encoding HumanCD40L TNF domain was expressed in Escherichia Coli.
Recombinant CD40 Ligand is a monomer protein consisting of150 amino acid residue subunits, and migrates as an approximately16 kDa protein under non-reducing and reducing conditions in SDS-PAGE.
>95%, as determined by SDS-PAGE and HPLC
The ED(50) was determined by the dose-dependentstimulation of IL-8 production by human PBMC and was found to be in the range of.0-20.0 ng/ml.
MIETYNQTSP RSAATGLPIS MKIFMYLLTV FLITQMIGSA LFAVYLHRRL DKIEDERNLH EDFVFMKTIQ RCNTGERSLS LLNCEEIKSQ FEGFVKDIML NKEETKKENS FEMQKGDQNP QIAAHVISEA SSKTTSVLQW AEKGYYTMSN NLVTLENGKQ LTVKRQGLYY IYAQVTFCSN REASSQAPFI ASLCLKSPGR FERILLRAAN THSSAKPCGQ QSIHLGGVFE LQPGASVFVN VTDPSQVSHG TGFTSFGLLK L
Endotoxin content was assayed using a LAL gel clot method. Endotoxin level was found to be less than.1 ng/µg(1EU/µg).
CD40 ligandwas lyophilized from a.2 μm filtered PBS solution pH7.0.
A quick spin of the vial followed by reconstitution in distilled water to a concentration not less than.1 mg/mL. This solution can then be diluted into other buffers.
The lyophilized protein is stable for at least years from date of receipt at -20° C. Upon reconstitution, this cytokine can be stored in working aliquots at° -° C for one month, or at -20° C for six months, with a carrier protein without detectable loss of activity. Avoid repeated freeze/thaw cycles.
This cytokine product is for research purposes only.It may not be used for therapeutics or diagnostic purposes.
The co-culture of primary CLL cells with BMSC, CD40L and CpG ODN promotes an immunophenotype comparable to that from proliferating CLL cells found in PB (A) PBMC from 40 patients diagnosed with CLL were used to analyze by FC the differential expression of CD38, CD49d, CD62L, CXCR4, CXCR5 and CCR7 in Ki-67-negative vs. positive CLL cells.
- Primary CLL cells from 12 patients were cultured in suspension or in co-culture with BMSC, CD40L and CpG ODN for 48 hours and the expression ofCD38, CD49d, CD62L, CXCR4, CXCR5 and CCR7 were analyzed.