VISTA/B7-H5/PD-1H Antibody (PSH0-65)

Novus Biologicals | Catalog # NBP3-33095

Recombinant Monoclonal Antibody
Novus Biologicals
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Key Product Details

Species Reactivity

Human

Applications

Immunohistochemistry, Immunohistochemistry-Paraffin, Western Blot, Flow Cytometry, Immunocytochemistry/ Immunofluorescence

Label

Unconjugated

Antibody Source

Recombinant Monoclonal Rabbit IgG Clone # PSH0-65 expressed in HEK293
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Product Specifications

Immunogen

Recombinant protein within human VISTA/B7-H5/PD-1H aa 1-194 / 311. (Uniprot: Q9H7M9)

Localization

Cell membrane.

Clonality

Monoclonal

Host

Rabbit

Isotype

IgG

Theoretical MW

34 kDa.
Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.

Scientific Data Images for VISTA/B7-H5/PD-1H Antibody (PSH0-65)

VISTA/B7-H5/PD-1H Antibody (PSH0-65)

Western Blot: VISTA/B7-H5/PD-1H Antibody (PSH0-65) [NBP3-33095]

Western Blot: VISTA/B7-H5/PD-1H Antibody (PSH0-65) [NBP3-33095] - Western blot analysis of VISTA/B7-H5/PD-1H on human lung tissue lysates with Rabbit anti-VISTA/B7-H5/PD-1H antibody (NBP3-33095) at 1/1,000 dilution.

Lysates/proteins at 40 ug/Lane.

Predicted band size: 34 kDa
Observed band size: 55 kDa

Exposure time: 30 seconds;

10% 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 (NBP3-33095) at 1/1,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1:100,000 dilution was used for 1 hour at room temperature.
VISTA/B7-H5/PD-1H Antibody (PSH0-65)

Immunohistochemistry: VISTA/B7-H5/PD-1H Antibody (PSH0-65) [NBP3-33095]

Immunohistochemistry: VISTA/B7-H5/PD-1H Antibody (PSH0-65) [NBP3-33095] - Immunohistochemical analysis of paraffin-embedded human brain tissue with Rabbit anti-VISTA/B7-H5/PD-1H antibody (NBP3-33095) at 1/200 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 (NBP3-33095) at 1/200 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.
VISTA/B7-H5/PD-1H Antibody (PSH0-65)

Immunocytochemistry/ Immunofluorescence: VISTA/B7-H5/PD-1H Antibody (PSH0-65) [NBP3-33095]

Immunocytochemistry/ Immunofluorescence: VISTA/B7-H5/PD-1H Antibody (PSH0-65) [NBP3-33095] - Immunocytochemistry analysis of PBMC labeling VISTA/B7-H5/PD-1H with Rabbit anti-VISTA/B7-H5/PD-1H antibody (NBP3-33095) at 1/100 dilution.

Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, permeabilized with 0.05% Triton X-100 in PBS for 20 minutes, and then blocked with 2% negative goat serum for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-VISTA/B7-H5/PD-1H antibody (NBP3-33095) at 1/100 dilution in 2% negative goat serum overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488) was used as the secondary antibody at 1/1,000 dilution. Nuclear DNA was labelled in blue with DAPI.

Beta tubulin (red) was stained at 1/200 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594) was used as the secondary antibody at 1/1,000 dilution.

Applications for VISTA/B7-H5/PD-1H Antibody (PSH0-65)

Application
Recommended Usage

Flow Cytometry

1:500-1:1000

Immunocytochemistry/ Immunofluorescence

1:100

Immunohistochemistry-Paraffin

1:200-1:1000

Western Blot

1:1000

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Formulation, Preparation, and Storage

Purification

Protein A purified

Formulation

PBS (pH7.4), 0.1% BSA and 40% Glycerol

Preservative

0.05% Sodium Azide

Concentration

1 mg/ml

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.

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

VISTA (V-domain immunoglobulin suppressor of T cell activation) is a cell surface coinhibitory protein belonging to the CD28/B7 family. VISTA can act as both a ligand and a receptor in regulating T cell activity. VISTA is expressed on dendritic cells, monocytes, and both CD4+ and CD8+ T cells. VISTA functions as an inhibitory receptor downregulating CD4+ and CD8+ T cell proliferation and exhibiting a protective effect in mice from graft versus host disease, asthma, and acute hepatitis. VISTA also acts as a ligand on antigen presenting cells. Two cell surface expressed proteins VSIG-3 and PSGL-1 have been identified as binding partners of VISTA. Immune checkpoint proteins like the VISTA protein are often overexpressed by cancer cells or surrounding cells in the tumor microenvironment. VISTA upregulation occurs on tumor-infiltrating leukocytes and is expressed on several cancer types including melanoma, lung, ovarian, colon, prostate, and pancreatic cancer. Further, knockout and blocking antibody studies of VISTA have shown an increase in T cell responses in several mouse tumor models making VISTA a potential therapeutic target alone or in combination with other known immune checkpoint inhibitors. Alternative names of VISTA include PD-1H, Gi24, Dies1, and SISP1.

Alternate Names

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

Gene Symbol

VSIR

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

Product Documents for VISTA/B7-H5/PD-1H Antibody (PSH0-65)

Certificate of Analysis

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Product Specific Notices for VISTA/B7-H5/PD-1H Antibody (PSH0-65)

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.

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Protocols

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