VISTA/B7-H5/PD-1H Antibody [Unconjugated]

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.

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.

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).

• Reactivity: Hu
• Applications: WB, ELISA, Flow, CyTOF-ready
• Clonality: Polyclonal
• Host: Sheep
• Conjugate: Unconjugated

VISTA/B7-H5/PD-1H Antibody [Unconjugated] Summary

• Immunogen: Mouse myeloma cell line NS0-derived recombinant human VISTA/B7-H5/PD-1H
  Phe33-Ala194
  Accession # Q9H7M9
• Specificity: Detects human VISTA/B7-H5/PD-1H in direct ELISAs and Western blots.
• Source: N/A
• Isotype: IgG
• Clonality: Polyclonal
• Host: Sheep
• Purity Statement: Antigen Affinity-purified

Applications/Dilutions

• Dilutions:
  • CyTOF-ready
  • ELISA
  • Flow Cytometry 0.25 ug/10^6 cells
  • Western Blot 1 ug/mL

Packaging, Storage & Formulations

• 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.
• Buffer: Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.
• Preservative: No Preservative
• Reconstitution Instructions: Reconstitute at 0.2 mg/mL in sterile PBS.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for VISTA/B7-H5/PD-1H Antibody [Unconjugated]

• 4632428N05Rik
• B7H5
• B7-H5
• C10orf54
• chromosome 10 open reading frame 54
• Dies1
• Gi24
• PD1H
• PD-1H
• platelet receptor Gi24
• PP2135
• SISP1
• stress induced secreted protein 1
• VISTA
• VSIR

Background

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.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for VISTA/B7-H5/PD-1H Antibody (AF7126)(2)

Publications using AF7126

• Fu, J;Li, S;Ma, H;Yang, J;Pagnotti, GM;Brown, LM;Weiss, SJ;Mapara, MY;Lentzsch, S; The checkpoint inhibitor PD-1H/VISTA controls osteoclast-mediated multiple myeloma bone disease Nature communications 2023-07-17 [PMID: 37460553] (Western Blot, Human)
• Wang J, Wu G, Manick B, Hernandez V, Renelt M, Erickson C, Guan J, Singh R, Rollins S, Solorz A, Bi M, Li J, Grabowski D, Dirkx J, Tracy C, Stuart T, Ellinghuysen C, Desmond D, Foster C, Kalabokis V VSIG-3 as a ligand of VISTA inhibits human T-cell function. Immunology, 2018-10-10;156(1):74-85. 2018-10-10 [PMID: 30220083] (Neutralization, Human)

Bioinformatics

• Uniprot:
  • Q9H7M9(Human)