Human VISTA/B7-H5/PD-1H Antibody Summary
Accession # Q9H7M9
This antibody functions as an ELISA detection antibody when paired with Mouse Anti-Human VISTA/B7-H5/PD-1H Monoclonal Antibody (Catalog # MAB71264).
This product is intended for assay development on various assay platforms requiring antibody pairs. We recommend the Human VISTA/B7-H5/PD-1H DuoSet ELISA Kit (Catalog # DY7126-05) for convenient development of a sandwich ELISA.
Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.
Detection of Human VISTA/B7-H5/PD-1H by Western Blot. Western blot shows lysates of human placenta tissue. PVDF membrane was probed with 1 µg/mL of Sheep Anti-Human VISTA/B7‑H5/PD‑1H Antigen Affinity-purified Polyclonal Antibody (Catalog # AF7126) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). A specific band was detected for VISTA/B7-H5/PD-1H at approximately 45-65 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Detection of VISTA/B7-H5/PD-1H in Human PBMCs by Flow Cytometry. Human peripheral blood mononuclear cells (PBMCs) were stained with Mouse Anti-Human CD11b/Integrin alpha M APC‑conjugated Monoclonal Antibody (Catalog # FAB16991A) and either (A) Sheep Anti-Human VISTA/B7-H5/PD-1H Antigen Affinity-purified Polyclonal Antibody (Catalog # AF7126) or (B) Normal Sheep IgG Control (Catalog # 5-001-A) followed by Phycoerythrin-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # F0126). View our protocol for Staining Membrane-associated Proteins.
Human VISTA/B7-H5/PD-1H ELISA Standard Curve. Recombinant Human VISTA/B7‑H5/PD‑1H protein was serially diluted 2-fold and captured by Mouse Anti-Human VISTA/B7‑H5/PD‑1H Monoclonal Antibody (Catalog # MAB71264) coated on a Clear Polystyrene Microplate (Catalog # DY990). Sheep Anti-Human VISTA/B7-H5/PD-1H Antigen Affinity-purified Polyclonal Antibody (Catalog # AF7126) was biotinylated and incubated with the protein captured on the plate. Detection of the standard curve was achieved by incubating Streptavidin-HRP (Catalog # DY998) followed by Substrate Solution (Catalog # DY999) and stopping the enzymatic reaction with Stop Solution (Catalog # DY994).
Preparation and Storage
- 12 months from date of receipt, -20 to -70 °C as supplied.
- 1 month, 2 to 8 °C under sterile conditions after reconstitution.
- 6 months, -20 to -70 °C under sterile conditions after reconstitution.
Platelet Receptor Gi24, also known as VISTA (V-domain Ig suppressor of T cell activation), B7-H5, B7H5, Dies1 (Differentiation of ESC-1), SISP1 and C10orf54, is a 55-65 kDa member of the Ig superfamily. It is a transmembrane molecule expressed in bone, on embryonic stem cells (ESCs), and on tumor cell surfaces. On ESCs, VISTA appears to positively interact with BMP-4, potentiating BMP signaling and the transition from an undifferentiated to a differentiated state. On tumor cells, VISTA both promotes MT1-MMP expression and activity and serves as a substrate for MT1-MMP. This increases the potential for cell motility. Mature human VISTA contains a 162 aa extracellular region with one V-type Ig-like domain and a 96 aa cytoplasmic domain. Human VISTA undergoes proteolytic cleavage by MT1-MMP, generating a soluble 30 kDa extracellular fragment plus a 25-30 kDa membrane-bound fragment. Over aa 33-194, human VISTA shares 70% and 67% aa identity with mouse and rat Gi24, respectively.
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