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 (1). Polymorphism of the CD272 molecule has been linked to rheumatoid arthritis (3). Applications: Suitable for use in ELISA and Flow Cytometry. Other applications not tested. Recommended Dilution: Flow Cytometry: 10ug/ml, Optimal dilutions to be determined by the researcher. Flow Cytometry Protocol: 1. Ficoll prepared human PBMC cells per tube were pelleted and pre-incubated for 5 minutes on ice with 20ul of human IgG at 300ug/ml. , 2. Cells were washed and incubated for 45 minutes on ice with 80ul of B5001-61J at 10ug/ml, 3. Cells were washed twice and incubated with 2º reagent Streptavidin/R-PE (S7973-88P), after which they were washed three times, fixed and analyzed using FACS. , 4. Cells stained positive with a mean shift of 1.32 log10 fluorescent units when compared to a Mouse IgG1/Biotin negative control at a similar concentration. , 5. Binding was blocked when cells were pre-incubated for 10 minuted with 20ul of 0.5mg/ml unlabeled anti-CD272 (B5001-61G). Storage and Stability: Store product at 4°C if to be used immediately within two weeks. For long-term storage, aliquot to avoid repeated freezing and thawing and store at -20°C. Aliquots are stable at -20°C for 12 months after receipt. Dilute required amount only prior to immediate use. Further dilutions can be made in assay buffer. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. ,