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VISTA Recombinant Rabbit Monoclonal Antibody [PSH0-64] - BSA and Azide free

Usd: 649

Specification

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Catalog# HA750641

VISTA Recombinant Rabbit Monoclonal Antibody [PSH0-64] - BSA and Azide free

Application

  • WB

  • IHC-P

  • IF-Tissue

Reactivity

  • Human

With BSA and Azide
Conjugation

  • unconjugated

Overview

Product Name

VISTA Recombinant Rabbit Monoclonal Antibody [PSH0-64] - BSA and Azide free

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Recombinant protein within human VISTA aa 1-194 / 311.

Species Reactivity

Human

Validated Applications

WB, IHC-P, IF-Tissue

Target Molecular Weight

Predicted band size: 34 kDa

Positive Control

A431 cell lysate, A375 cell lysate, K-562 cell lysate, TF-1 cell lysate, human lung tissue lysate, human brain tissue, human placenta tissue, human tonsil tissue.

Conjugation

unconjugated

Clone Number

PSH0-64

Product Features

Form

Liquid

Concentration

1mg/ml

Storage Instructions

Store at 2-8℃. Avoid freeze.

Storage Buffer

1*PBS (pH7.4).

Isotype

IgG

Purification Method

Protein A affinity purified.

Application Dilution

Target

Function

V-domain Ig suppressor of T cell activation (VISTA) is a type I transmembrane protein that functions as an immune checkpoint and is encoded by the C10orf54 gene. VISTA is approximately 50kDa and belongs to the immunoglobulin superfamily and has one IgV domain. VISTA is part of the B7 family, is primarily expressed in white blood cells and its transcription is partially controlled by p53. There is evidence that VISTA can act as both a ligand and a receptor on T cells to inhibit T cell effector function and maintain peripheral tolerance. VISTA is produced at high levels in tumor-infiltrating lymphocytes, such as myeloid-derived suppressor cells and regulatory T cells, and its blockade with an antibody results in delayed tumor growth in mouse models of melanoma and squamous cell carcinoma. Monocytes from HIV-infected patients produce higher levels of VISTA compared to uninfected individuals. The increased VISTA levels correlated with an increase in immune activation and a decrease in CD4-positive T cells. There is an ongoing cancer immunotherapy clinical trial for a monoclonal antibody targeting VISTA in advanced cancer. Preliminary results of the phase I clinical trial show good safety tolerance and anti-cancer activity in patients with advanced tumours. Another ongoing clinical trial involves a small molecule that antagonizes the programmed death-ligands 1 and 2 (PD-L1 and PD-L2), and VISTA pathways in patients with advanced solid tumors or lymphomas.

Background References

1. Yuan L et al. VISTA: A Mediator of Quiescence and a Promising Target in Cancer Immunotherapy. Trends Immunol. 2021 Mar

2. Huang X et al. VISTA: an immune regulatory protein checking tumor and immune cells in cancer immunotherapy. J Hematol Oncol. 2020 Jun

Subcellular Location

Cell membrane.

UNIPROT

SwissProt: Q9H7M9 Human

Synonyms

B7 H5 antibody

B7H5 antibody

C10orf54 antibody

chromosome 10 open reading frame 54 antibody

DD1alpha antibody

GI24 antibody

GI24_HUMAN antibody

PD-1H antibody

PDCD1 homolog antibody

Platelet receptor Gi24 antibody

Expand

Images

  • <span style="font-weight: bold;">☑ Relative expression (RE)</span><br /><br />This data was developed using <a href="/products/HA721452" style="font-weight: bold;text-decoration: underline;">HA721452</a>, the same antibody clone in a different buffer formulation.<br />Western blot analysis of VISTA on different lysates with Rabbit anti-VISTA antibody (<a href="/products/HA721452" style="font-weight: bold;text-decoration: underline;">HA721452</a>) at 1/1,000 dilution and competitor's antibody at 1/1,000 dilution.<br /><br />Lane 1: A431 cell lysate (20 µg/Lane)<br />Lane 2: A375 cell lysate (20 µg/Lane)<br />Lane 3: K-562 cell lysate (20 µg/Lane)<br />Lane 4: TF-1 cell lysate (20 µg/Lane)<br />Lane 5: MCF7 cell lysate (low expression) (20 µg/Lane)<br />Lane 6: HEK-293 cell lysate (low expression) (20 µg/Lane)<br />Lane 7: Human lung tissue lysate (30 µg/Lane)<br /><br />Predicted band size: 34 kDa<br />Observed band size: 45-60 kDa (Glycosylated)<br /><br />Exposure time: 30 seconds; ECL: K1801;<br /><br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/HA721452" style="font-weight: bold;text-decoration: underline;">HA721452</a>) at 1/1,000 dilution and competitor's antibody at 1/1,000 dilution were used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1/50,000 dilution was used for 1 hour at room temperature.

    ☑ Relative expression (RE)

    This data was developed using HA721452, the same antibody clone in a different buffer formulation.
    Western blot analysis of VISTA on different lysates with Rabbit anti-VISTA antibody (HA721452) at 1/1,000 dilution and competitor's antibody at 1/1,000 dilution.

    Lane 1: A431 cell lysate (20 µg/Lane)
    Lane 2: A375 cell lysate (20 µg/Lane)
    Lane 3: K-562 cell lysate (20 µg/Lane)
    Lane 4: TF-1 cell lysate (20 µg/Lane)
    Lane 5: MCF7 cell lysate (low expression) (20 µg/Lane)
    Lane 6: HEK-293 cell lysate (low expression) (20 µg/Lane)
    Lane 7: Human lung tissue lysate (30 µg/Lane)

    Predicted band size: 34 kDa
    Observed band size: 45-60 kDa (Glycosylated)

    Exposure time: 30 seconds; ECL: K1801;

    4-20% SDS-PAGE gel.

    Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA721452) at 1/1,000 dilution and competitor's antibody at 1/1,000 dilution were used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

  • <span style="font-weight: bold;">☑ Knockdown (KD)</span><br /><br />This data was developed using <a href="/products/HA721452" style="font-weight: bold;text-decoration: underline;">HA721452</a>, the same antibody clone in a different buffer formulation.<br />Western blot analysis of VISTA on different lysates with Rabbit anti-VISTA antibody (<a href="/products/HA721452" style="font-weight: bold;text-decoration: underline;">HA721452</a>) at 1/1,000 dilution.<br /><br />Lane 1: A431-si NT cell lysate<br />Lane 2: A431-si VISTA cell lysate<br /><br />Lysates/proteins at 10 µg/Lane.<br /><br />Predicted band size: 34 kDa<br />Observed band size: 45-60 kDa (Glycosylated)<br /><br />Exposure time: 28 seconds; ECL: K1801;<br /><br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/HA721452" style="font-weight: bold;text-decoration: underline;">HA721452</a>) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1/50,000 dilution was used for 1 hour at room temperature.

    ☑ Knockdown (KD)

    This data was developed using HA721452, the same antibody clone in a different buffer formulation.
    Western blot analysis of VISTA on different lysates with Rabbit anti-VISTA antibody (HA721452) at 1/1,000 dilution.

    Lane 1: A431-si NT cell lysate
    Lane 2: A431-si VISTA cell lysate

    Lysates/proteins at 10 µg/Lane.

    Predicted band size: 34 kDa
    Observed band size: 45-60 kDa (Glycosylated)

    Exposure time: 28 seconds; ECL: K1801;

    4-20% SDS-PAGE gel.

    Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA721452) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

  • <span style="font-weight: bold;">☑ Cell treatment (CT)</span><br /><br />This data was developed using <a href="/products/HA721452" style="font-weight: bold;text-decoration: underline;">HA721452</a>, the same antibody clone in a different buffer formulation.<br />Western blot analysis of VISTA on different lysates with Rabbit anti-VISTA antibody (<a href="/products/HA721452" style="font-weight: bold;text-decoration: underline;">HA721452</a>) at 1/1,000 dilution.<br /><br />Lane 1: THP-1 cell lysate<br />Lane 2: THP-1 cell lysate treated with deglycosylation<br /><br />Lysates/proteins at 20 µg/Lane.<br /><br />Predicted band size: 34 kDa<br />Observed band size: 34/45-60 kDa (Glycosylated)<br /><br />Exposure time: 40 seconds; ECL: K1801;<br /><br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/HA721452" style="font-weight: bold;text-decoration: underline;">HA721452</a>) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1/50,000 dilution was used for 1 hour at room temperature.

    ☑ Cell treatment (CT)

    This data was developed using HA721452, the same antibody clone in a different buffer formulation.
    Western blot analysis of VISTA on different lysates with Rabbit anti-VISTA antibody (HA721452) at 1/1,000 dilution.

    Lane 1: THP-1 cell lysate
    Lane 2: THP-1 cell lysate treated with deglycosylation

    Lysates/proteins at 20 µg/Lane.

    Predicted band size: 34 kDa
    Observed band size: 34/45-60 kDa (Glycosylated)

    Exposure time: 40 seconds; ECL: K1801;

    4-20% SDS-PAGE gel.

    Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA721452) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

  • This data was developed using <a href="/products/HA721452" style="font-weight: bold;text-decoration: underline;">HA721452</a>, the same antibody clone in a different buffer formulation.<br />Immunohistochemical analysis of paraffin-embedded human brain tissue with Rabbit anti-VISTA antibody (<a href="/products/HA721452" style="font-weight: bold;text-decoration: underline;">HA721452</a>) at 1/1,000 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/HA721452" style="font-weight: bold;text-decoration: underline;">HA721452</a>) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    This data was developed using HA721452, the same antibody clone in a different buffer formulation.
    Immunohistochemical analysis of paraffin-embedded human brain tissue with Rabbit anti-VISTA antibody (HA721452) at 1/1,000 dilution.

    The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA721452) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • This data was developed using <a href="/products/HA721452" style="font-weight: bold;text-decoration: underline;">HA721452</a>, the same antibody clone in a different buffer formulation.<br />Immunohistochemical analysis of paraffin-embedded human placenta tissue with Rabbit anti-VISTA antibody (<a href="/products/HA721452" style="font-weight: bold;text-decoration: underline;">HA721452</a>) at 1/1,000 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/HA721452" style="font-weight: bold;text-decoration: underline;">HA721452</a>) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    This data was developed using HA721452, the same antibody clone in a different buffer formulation.
    Immunohistochemical analysis of paraffin-embedded human placenta tissue with Rabbit anti-VISTA antibody (HA721452) at 1/1,000 dilution.

    The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA721452) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • This data was developed using <a href="/products/HA721452" style="font-weight: bold;text-decoration: underline;">HA721452</a>, the same antibody clone in a different buffer formulation.<br />Immunohistochemical analysis of paraffin-embedded human tonsil tissue with Rabbit anti-VISTA antibody (<a href="/products/HA721452" style="font-weight: bold;text-decoration: underline;">HA721452</a>) at 1/1,000 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/HA721452" style="font-weight: bold;text-decoration: underline;">HA721452</a>) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    This data was developed using HA721452, the same antibody clone in a different buffer formulation.
    Immunohistochemical analysis of paraffin-embedded human tonsil tissue with Rabbit anti-VISTA antibody (HA721452) at 1/1,000 dilution.

    The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA721452) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • This data was developed using <a href="/products/HA721452" style="font-weight: bold;text-decoration: underline;">HA721452</a>, the same antibody clone in a different buffer formulation.<br />Application: IF-Tissue<br /><br />Species: Human<br /><br />Site: tonsil<br /><br />Sample: Paraffin-embedded section<br /><br />Antibody concentration: 1/500

    This data was developed using HA721452, the same antibody clone in a different buffer formulation.
    Application: IF-Tissue

    Species: Human

    Site: tonsil

    Sample: Paraffin-embedded section

    Antibody concentration: 1/500

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