Anti-BTLA (B- and T-lymphocyte Associated Protein, B- and T-lymphocyte Attenuator, BTLA1, CD272, CD2

Human CD272 (BTLA, B and T Lymphocyte Attenuator) is a member of the Immunoglobulin superfamily and has sequence homology to PD-1 and CTLA4. It is expressed on T and B lymphocytes and other hemopoetic lineages. Engagement of this molecule by its ligand, CD270 (HVEM, a TNF superfamily member) can down regulate activated T and B cell responses. CD272 levels on antigen specific CD8+ T cells have been reported to decrease in viral specific, but not melanoma specific activated lines. Polymorphism of the CD272 molecule has been linked to rheumatoid arthritis. Applications: Suitable for use in FLISA and Flow Cytometry. Other applications not tested. Recommended Dilutions: FLISA: Detection, Flow Cytometry: 1:50, Optimal dilutions to be determined by the researcher. Flow Cytometry Protocol: 1. 5x10e5 ficoll prepared human peripheral blood mononuclear cells (PBMC) per tube were pelleted and pre incubated 5 minutes with 20ul of human IgG at 300ug/ml, after which they were washed and incubated 45 minutes on ice with 80ul of B5001-61K at a 1:50 dilution (5ug/ml). , 2. They were then washed three times, fixed and analyzed by FACS using a lymphoid gate. , 3. Cells stained positive with a mean shift of 0.54 log10 fluorescent units when compared to a Mouse IgG1 (FITC) negative control at a similar concentration. , 4. Binding was blocked when cells were pre incubated 10 minutes with 20ul of 0.5mg/ml of B5001-61G. Storage and Stability: May be stored at 4°C for short-term only. Aliquot to avoid repeated freezing and thawing. Store at -20°C. Aliquots are stable for 12 months. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. FITC conjugates are sensitive to light.