CD154 protein

CD154-muCD8-Biotin

Cat. Num. AAA666788

• Reactivity: Human, Pig, Dog, Mouse
• Applications: Flow Cytometry, Functional Assay, ELISA, Western Blot
• Synonyms: CD154; CD154-muCD8-Biotin; Human CD154 (CD40 Ligand) muCD8/Biotin; CD154 protein

• Host: CHO cells
• Reactivity: Human, Pig, Dog, Mouse
• Form/Format: 50 mM Sodium Phosphate pH 7.5, 100 mM Potassium Chloride, 150mM NaCl, 5% Glycerol, 0.2% BSA.
• Concentration: 0.5 mg/ml (varies by lot)

• Applicable Applications for CD154 protein: Flow Cytometry (FC/FACS), ELISA (EIA), Western Blot (WB)
• Molecular Structure: A soluble fusion protein consisting of the extracellular domain (213aa) of human CD154 fused to the extracellular domain (167aa) of murine CD8 alpha.
• Transfectant Cell Line: CHO
• Performance: Five x 105 cultured human Raji cells per tube were washed and incubated 45 minutes on ice with 80 ul of CD154-muCD8/Biotin at a concentration of 10 ug/ml. Cells were washed twice and incubated with 2 degree reagent Streptavidin/R-PE, after which they were washed three times, fixed and analyzed by FACS. Cells stained positive with a mean shift of 1.28 log10 fluorescent units when compared to a buffer control at a similar concentration. Binding was blocked when reagent was pre incubated 30 minutes with anti-CD154 antibody at a concentration of 100 ug/ml.
• Conjugation: Biotin
• Production: Fusion protein from (low FBS containing) tissue culture supernatant of transfectants was purified using affinity and size exclusion chromatography), and reacted with NHS-Biotin. Unconjugated Biotin was removed from conjugate by desalting.
• Buffer: 50 mM Sodium Phosphate pH 7.5, 100 mM Potassium Chloride, 150mM NaCl, 5% Glycerol, 0.2% BSA, 0.04% NaN3 (as a preservative).
• Preparation and Storage: Store at 2 - 5 degree C. Freeze/Thawing is not recommended.; Product should retain activity for at least 6 months after shipping date when stored as recommended.

Testing Data

Related Product Information for CD154 protein

Information: Human CD154 (CD40 Ligand) is a member of the tumor necrosis factor (TNF) family and is expressed on the surface of activated T cells. Interaction of CD154 and CD40 is essential for isotype switching in B cells. Known genetic defects that alter this interaction lead to impaired immune system function (1). CD154 has been shown to be hyperexpressed by B and T cells in SLE patients (4). CD154 has been reported to be expressed on vascular endothelial cells, smooth muscle cells and macrophages indicating a possible role for the CD40-CD154 immunoregulatory signaling mechanism during inflammation and immunity in atherogenesis (6). Human CD154-muCD8 binds to cell surface expressed human CD40 and this binding is blocked by anti-human CD154 monoclonal antibody.

References

1) D. Gray, et al, (1994) Seminars in Immunol 6: 303-310. 2) F. Pietravalle, et al, (1996) J Biol Chemistry 271: 5965-5967. 3) R.J. Noelle, (1996) Immunity 4: 415-419. 4) A. Desai-Mehta, et al, (1996) J Clin Invest 97: 2063-2073. 5) I.S. Grewal and R.A. Flavell, (1996) Immunol Today 17: 410-414. 6) F. Mach, et al, (1997) Proc Natl Acad Sci USA 94:1931-1936.