VISTA Recombinant Rabbit Monoclonal Antibody [PSH0-64]
Catalog# HA721452
VISTA Recombinant Rabbit Monoclonal Antibody [PSH0-64]
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WB
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IHC-P
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IF-Tissue
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Human
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unconjugated
Overview
Product Name
VISTA Recombinant Rabbit Monoclonal Antibody [PSH0-64]
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
RRID
Reactivity Data
| WB | IHC-P | IF-Tissue | |
|---|---|---|---|
| Human |
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| Mouse |
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| Rat |
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Product Features
Form
Liquid
Concentration
1 mg/mL.(The concentration of this product may be batch-dependent)
Storage Instructions
Shipped at 4℃. Store at +4℃ short term (1-2 weeks). Store at -20℃ long term.
Storage Buffer
PBS (pH7.4), 0.1% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Isotype
IgG
Purification Method
Protein A affinity purified.
Application Dilution
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WB
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1:1,000
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IHC-P
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1:1,000
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IF-Tissue
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1:500
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
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
ExpandImages
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☑ Relative expression (RE)
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. -
☑ Knockdown (KD)
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. -
☑ Cell treatment (CT)
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. -
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. -
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. -
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. -
Application: IF-Tissue
Species: Human
Site: tonsil
Sample: Paraffin-embedded section
Antibody concentration: 1/500
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"