Key Product Details

Species Reactivity

Validated:

Human

Cited:

Human

Applications

Validated:

Western Blot, ELISA, Flow Cytometry, CyTOF-ready

Cited:

Western Blot, Neutralization

Label

Unconjugated

Antibody Source

Polyclonal Sheep IgG
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Product Specifications

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.

Clonality

Polyclonal

Host

Sheep

Isotype

IgG

Scientific Data Images for Human VISTA/B7-H5/PD-1H Antibody

Detection of Human VISTA/B7-H5/PD-1H antibody by Western Blot.

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 antibody in Human PBMCs antibody by Flow Cytometry.

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 Antibody in ELISA Standard Curve.

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).

Applications for Human VISTA/B7-H5/PD-1H Antibody

Application
Recommended Usage

CyTOF-ready

Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.

ELISA

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.

Flow Cytometry

0.25 µg/106 cells
Sample: Human peripheral blood mononuclear cells (PBMCs)

Western Blot

1 µg/mL
Sample: Human placenta tissue

Flow Cytometry Panel Builder

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Save time and reduce costly mistakes by quickly finding compatible reagents using the Panel Builder Tool.

Advanced Features

  • Spectra Viewer - Custom analysis of spectra from multiple fluorochromes
  • Spillover Popups - Visualize the spectra of individual fluorochromes
  • Antigen Density Selector - Match fluorochrome brightness with antigen density
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Formulation, Preparation, and Storage

Purification

Antigen Affinity-purified

Reconstitution

Reconstitute at 0.2 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.


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Formulation

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.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

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.

Calculators

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Background: VISTA/B7-H5/PD-1H

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.

Alternate Names

4632428N05Rik, B7-H5, C10orf54, Dies1, Gi24, PD-1H, PD1H, SISP1, VSIR

Entrez Gene IDs

64115 (Human); 74048 (Mouse); 690899 (Rat); 102116468 (Cynomolgus Monkey)

Gene Symbol

VSIR

UniProt

Additional VISTA/B7-H5/PD-1H Products

Product Documents for Human VISTA/B7-H5/PD-1H Antibody

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot or batch number in the search box below.

Note: Certificate of Analysis not available for kit components.

Product Specific Notices for Human VISTA/B7-H5/PD-1H Antibody

For research use only

Citations for Human VISTA/B7-H5/PD-1H Antibody

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Protocols

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