Staining of BALB/c splenocytes with Anti-Human/Mouse CD45R (B220) APC (cat. 17-0452) and 0.125 µg of Mouse IgG1 κ Isotype Control Purified (cat. 14-4714) (left) or 0.125 µg of Anti-Mouse CD272 (BTLA) Purified (right) followed by Anti-Mouse IgG Biotin (cat. 13-4013) and Streptavidin PE (cat. 12-4317). Total viable cells were used for analysis.
Flow Cytometry Product Notes: Test Sizes: To accommodate multicolor flow cytometry, eBioscience is in the process of reducing test size volumes from 20 µl to 5 µl. Please check your antibody vial for the recommended test size. Fluorochrome Replacements: eBioscience is in the process of replacing all Alexa Fluor® 647 conjugated products with eFluor® 660 conjugated products.
Description
The BTLA antibody reacts with mouse BTLA, B and T lymphocyte attenuator from both BALB/c and C57Bl/6 strains. BTLA is expressed by peripheral lymphocytes, splenic macrophages, developing B cells in the bone marrow and developing T cells in thymus and mature, but not immature bone marrow-derived dendritic cells. BTLA is implicated as a negative regulator of the activation and/or function of various hemopoietic cell types. It is reported that BTLA binds to B7-H4, but further studies are needed to confirm this interaction.
Note: The anti-mouse BTLA monoclonal antibody 6F7 is reported to stain CD4+ and CD8+ single-positive (SP) thymocytes (Hurchla et al). However, other anti-mouse BTLA clones generated simultaneously with 6F7 (8F4, 3F9.D12, 6G3 and 6H6) do not stain SP thymocytes . It is not understood why there is a discrepancy in thymocyte staining. However, in staining of splenocytes and bone marrow cells, clones 8F4, 3F9.D12, 6G3 and 6H6 stain similar populations to 6F7.
Applications Reported
For research use only, not for diagnostic or therapeutic use. This 8F4 antibody has been reported for use in flow cytometric analysis.
Applications Tested
This 8F4 antibody has been tested by flow cytometric analysis of mouse splenocytes. This can be used at less than or equal to 0.25 μg per test. A test is defined as the amount (μg) of antibody that will stain a cell sample in a final volume of 100 µL. Cell number should be determined empirically but can range from 105 to 108 cells/test. It is recommended that the antibody be carefully titrated for optimal performance in the assay of interest.
References
Watanabe N, Gavrieli M, Sedy JR, Yang J, Fallarino F, Loftin SK, Hurchla MA, Zimmerman N, Sim J, Zang X, Murphy TL, Russell JH, Allison JP, Murphy KM. 2003. BTLA is a lymphocyte inhibitory receptor with similarities to CTLA-4 and PD-1. Nat Immunol. 4 (7): 670-9.
Sedy JR, Gavrieli M, Potter KG, Hurchla MA, Lindsley RC, Hildner K, Scheu S, Pfeffer K, Ware CF, Murphy TL, Murphy KM. 2005. B and T lymphocyte attenuator regulates T cell activation through interaction with herpesvirus entry mediator. Nat Immunol. (6)1: 90-8.
Hurchla MA, Sedy JR, Gavrielli M, Drake CG, Murphy TL, Murphy KM. 2005. B and T Lymphocyte Attenuator exhibits structural expression polymorphisms and is highly induced in CD4+ T cells. The Journal of Immunology. 174: 3377-3385.
