The MIH65.rMAb is a recombinant monoclonal antibody derived from MIH65 hybridoma cells. MIH65.rMAb specifically binds to human V-domain Ig suppressor of T cell activation (VISTA) like the conventional MIH65 antibody and performs like the MIH65 antibody when used to stain cells and analyze them by flow cytometry. VISTA is also known as B7-H5 (B7H5), PD-1H, DD1alpha (DD1α), chromosome 10 open reading frame 54 (C10orf54), Platelet Receptor Gi24 (GI24), or Stress induced secreted protein 1 (SISP1). VISTA is a 55-65 kDa single-pass, type I transmembrane glycoprotein that is encoded by VSIR (V-set immunoregulatory receptor) which belongs to the V-set domain containing family within the Ig superfamily. VISTA is primarily expressed on precursors and mature cells of the hematopoietic lineage including, monocytes, macrophages, neutrophils, dendritic cells (DC), T cells, regulatory T cells (Treg), and natural killer (NK) cells but not B cells. VISTA functions as an inhibitory regulator that can suppress T cell proliferation and cytokine production.
The antibody was conjugated to BD Horizon BUV615 which is part of the BD Horizon Brilliant™ Ultraviolet family of dyes. This dye is a tandem fluorochrome with an Ex Max near 350 nm and an Em Max near 615 nm. BD Horizon Brilliant BUV615 can be excited by the ultraviolet laser (355 nm) and detected with a 610/20 filter and a 595 nm LP. Due to the excitation of the acceptor dye by the blue/yellow-green laser line, there may be significant spillover into channels detecting PE-CF594 like emissions (eg, 610/20-nm filter).