VISTA Monoclonal Antibody (B7H5DS8), PE-eFluor™ 610, eBioscience™, Invitrogen™

Description

Description: This B7H5DS8 monoclonal antibody recognizes human VISTA, also known as B7-H5 or PD-1H. This clone has been tested and shown not to cross-react with human B7-H4, B7-H6 and B7-H7. This B7H5DS8 antibody will work in flow cytometry only on live and not paraformaldehyde-fixed cells. Applications Reported: This B7H5DS8 antibody has been reported for use in flow cytometric analysis. Applications Tested: This B7H5DS8 antibody has been pre-diluted and tested by flow cytometric analysis of normal human peripheral blood cells. This may be used at 5 μL (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 10^5 to 10^8 cells/test. PE-eFluor 610 can be excited with laser lines from 488-561 nm and emits at 607 nm. We recommend using a 610/20 band pass filter (equivalent to PE-Texas Red). Please make sure that your instrument is capable of detecting this fluorochrome. Light sensitivity: This tandem dye is sensitive to photo-induced oxidation. Please protect this vial and stained samples from light. Fixation: Samples can be stored in IC Fixation Buffer (Product # 00-8222-49) (100 μL of cell sample + 100 μL of IC Fixation Buffer) or 1-step Fix/Lyse Solution (Product # 00-5333-57) for up to 3 days in the dark at 4°C with minimal impact on brightness and FRET efficiency/compensation. Some generalizations regarding fluorophore performance after fixation can be made, but clone specific performance should be determined empirically. Excitation: 488-561 nm; Emission: 607 nm; Laser: Blue Laser, Green Laser, Yellow-Green Laser.

VISTA (V-domain Ig containing suppressor of T-cell activation), also known as B7-H5 or PD-1H, is a product of the VSIR gene. Although VISTA bears sequence homology to PD-L1 it has a different expression pattern. In both mouse and humans, VISTA is expressed predominantly on hematopoietic cells with the highest densities on myeloid cells, and lower levels on T cells. It is usually absent on B cells. VISTA expressed by antigen presenting cells has been shown to directly suppress proliferation and cytokine production of CD4 and CD8 T cells, making it a target for cancer immunotherapy. The reported ligands of VISTA include CD28H, VSIG-3 and VSIG-8.