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 Integrin alpha M/CD11b PE-conjugated Monoclonal Antibody (Catalog # FAB16991P) and either (A) Mouse IgG2B Isotype Control (Catalog # MAB0041) or (B) Mouse Anti-Human VISTA/B7-H5/PD-1H Monoclonal Antibody (Catalog # MAB71261) followed by Allophycocyanin-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # F0101B).
VISTA/B7-H5/PD-1H in BG01V Human Embryonic Stem Cells.
VISTA/B7-H5/PD-1H was detected in immersion fixed BG01V human embryonic stem cells using Mouse Anti-Human VISTA/B7-H5/PD-1H Monoclonal Antibody (Catalog # MAB71261) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (red, upper panel; Catalog # NL007) and counterstained with DAPI (blue, lower panel). Specific staining was localized to cell surfaces and cytoplasm. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.
Preparation and Storage
Sterile PBS to a final concentration of 0.5 mg/mL.
Reconstitution Buffer Available
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
Stability & Storage
Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
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, Gi24 appears to positively interact with BMP-4, potentiating BMP signaling and the transition from an undifferentiated to a differentiated state. On tumor cells, Gi24 both promotes MT1-MMP expression and activity and serves as a substrate for MT1-MMP. This increases the potential for cell motility. Mature human Gi24 contains a 162 aa extracellular region with one V-type Ig-like domain and a 96 aa cytoplasmic domain. Human Gi24 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 Gi24 shares 70% and 67% aa identity with mouse and rat Gi24, respectively.
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We have 1 review tested in 1 application: Flow Cytometry.
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Other Experimental Details
Other Experimental Details
immunophenotyping by flow cytometry. The antibody worked exactly as described to identify VISTA+ cells