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

Catalog # Availability Size / Price Qty
AF7126-100
AF7126-SP
Detection  of Human VISTA/B7-H5/PD-1H by Western Blot.
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Human VISTA/B7-H5/PD-1H Antibody Summary

Species Reactivity
Human
Specificity
Detects human VISTA/B7-H5/PD-1H in direct ELISAs and Western blots.
Source
Polyclonal Sheep IgG
Purification
Antigen Affinity-purified
Immunogen
Mouse myeloma cell line NS0-derived recombinant human VISTA/B7-H5/PD-1H
Phe33-Ala194
Accession # Q9H7M9
Formulation
Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied as a 0.2 µm filtered solution in PBS.
Label
Unconjugated

Applications

Recommended Concentration
Sample
Western Blot
1 µg/mL
See below
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
See below
CyTOF-ready
Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.
 

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.

Data Examples

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

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.

Flow Cytometry Detection of  VISTA/B7-H5/PD-1H in Human PBMCs by Flow Cytometry. View Larger

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).

Reconstitution Calculator

Reconstitution Calculator

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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Preparation and Storage

Reconstitution
Reconstitute at 0.2 mg/mL in sterile PBS.
Reconstitution Buffer Available
Reconstitution Buffer 1 (PBS)
Catalog #
Availability
Size / Price
Qty
RB01
Shipping
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.

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.

Entrez Gene IDs
64115 (Human); 74048 (Mouse); 690899 (Rat); 102116468 (Cynomolgus Monkey)
Alternate Names
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

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