VISTA Monoclonal Antibody (MH5A), PerCP-eFluor™ 710, eBioscience™, Invitrogen™

Description

VISTA is a homolog of PD-1 (PD-1H). PD-1H is a 309 aa type I transmembrane protein that belongs to the CD28 superfamily. This cell surface receptor is expressed on nearly all hematopoietic cells except B cells. VISTA negatively regulates T cell responses by suppressing early TCR activation, inhibiting cell proliferation, and suppressing TH1 cytokine production.

Description: The monoclonal antibody MH5A reacts with a homolog of PD-1 (PD-1H) also known as VISTA. PD-1H is a 309 aa type I transmembrane protein that belongs to the CD28 superfamily. This cell surface receptor is expressed on nearly all hematopoietic cells except B cells. VISTA negatively regulates T cell responses by suppressing early TCR activation, inhibiting cell proliferation, and suppressing TH1 cytokine production.The MH5A antibody has been reported to inhibit T cell infiltration in graft-versus-host disease mouse models.Applications Reported: This MH5A antibody has been reported for use in flow cytometric analysis.Applications Tested: This MH5A antibody has been tested by flow cytometric analysis of mouse splenocytes. This can be used at less than or equal to 1 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. It is recommended that the antibody be carefully titrated for optimal performance in the assay of interest.PerCP-eFluor™ 710 emits at 710 nm and is excited with the blue laser (488 nm); it can be used in place of PerCP-Cyanine5.5. We recommend using a 710/50 bandpass filter, however, the 695/40 bandpass filter is an acceptable alternative. Please make sure that your instrument is capable of detecting this fluorochrome.Fixation: Samples can be stored in IC Fixation Buffer (cat. 00-8222) (100 μL of cell sample + 100 μL of IC Fixation Buffer) or 1-step Fix/Lyse Solution (cat. 00-5333) 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 nm; Emission: 710 nm; Laser: Blue Laser.Filtration: 0.2μm post-manufacturing filtered.