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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 (Product # 00-8222) (100 µL of cell sample + 100 µL of IC Fixation Buffer) or 1-step Fix/Lyse Solution (Product # 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: 0.2 µm post-manufacturing filtered.
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.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Watch the video to learn how to use the Invitrogen Flow Cytometry Panel Builder to build your next flow cytometry panel in 5 easy steps.
Protein Aliases: differentiation of ESCs 1; PD-H1; Platelet receptor Gi24; V-set domain-containing immunoregulatory receptor; V-set immunoregulatory receptor; V-type immunoglobulin domain-containing suppressor of T-cell activation
Gene Aliases: Dies1; PD-1H; VISTA; Vsir
UniProt ID: (Mouse) Q9D659
Entrez Gene ID: (Mouse) 74048
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