Mouse Anti-BTLA (B- and T-lymphocyte Associated Protein, B- and T-lymphocyte Attenuator, BTLA1, CD272, CD272 Antigen, FLJ16065, MGC129743) (PE)
B- and T-lymphocyte Associated
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 FLISA and Flow Cytometry. Other applications not tested.
Recommended Dilution
Flow Cytometry: 1:50 (10ug/ml) 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 B5001-61L diluted 1:50 (10ug/ml). 2. Cells were washed three times, fixed and analyzed by FACS using a lymphoid gate. 3. Cells stained positive with a mean shift of 1.15 log10 fluorescent units when compared to a Mouse IgG1/R-PE negative control at a similar concentration. 4. Binding was blocked when cells were pre incubated 10 minutes with 20ul of 0.5mg/ml anti-CD272 antibody (Catalog #B5001-61G).
Storage and Stability
May be stored at 4°C before opening. DO NOT FREEZE! Stable at 4°C as an undiluted liquid. Dilute only prior to immediate use. Stable for 12 months at 4°C. Freezing R-Phycoerythrin (PE) conjugates will result in a substantial loss of activity. PE conjugates are sensitive to light.
Immunogen
Recombinant soluble human CD272.
Form
Supplied as a liquid in 50mM sodium phosphate pH 7.5, 500mM potassium chloride, 150mM sodium chloride, 0.2 BSA, 15% glycerol, 0.04% sodium azide. Labeled with R-Phycoerythrin (PE).
Purity
Supplied as a liquid in PBS, pH 7.5, 0.2% BSA, 0.04% sodium azide, 15% glycerol. Labeled with R-Phycoerythrin (PE).
Specificity
Recognizes human BTLA. Binds to CD272 on the cell surface of human PBMC in Flow cytometry and to recombinant CD272-muIg (B5001-61M) in EIA. Does not block binding of Biotinylated CD270(HVEM)-muIg (H2033-98A) to CD272-muIg in EIA, but appears to enhance this interaction.
