VISTA/B7-H5/PD-1H Antibody (BLR035F)

VISTA was detected in immersion fixed paraffin-embedded sections of human Spleen using Rabbit Anti-Human VISTA Recombinant Monoclonal Antibody (Catalog # NBP2-76406) at 1:250 at 37 ° Celsius for 4 minutes. Before incubation with the primary antibody, tissue underwent an all-in-one dewaxing and antigen retrieval preprocessing using PreTreatment Module (PT Module) and Dewax and HIER Buffer H (pH 9; Epredia Catalog # TA-999-DHBH). Tissue was stained using the Alexa Fluor™ Plus 647 Goat anti-Rabbit IgG Secondary Antibody at 1:200 at 37 ° Celsius for 2 minutes. (Yellow; Lunaphore Catalog # DR647RB) and counterstained with DAPI (blue; Lunaphore Catalog # DR100). Specific staining was localized to the membrane. Protocol available in COMET™ Panel Builder.

Western Blot: VISTA/B7-H5/PD-1H Antibody (BLR035F) [NBP2-76406] - Detection of human VISTA by WB of Jurkat, MOLT-4, MJ, RPMI-8226, HeLa, KG-1, HEK293T, and SUP-M2 lysate. Antibody: Rabbit anti-VISTA recombinant monoclonal [BLR035F] (NBP2-76406). Secondary: HRP-conjugated goat anti-rabbit IgG.Lower Panel: Rabbit anti-GAPDH.

Immunocytochemistry/Immunofluorescence: VISTA/B7-H5/PD-1H Antibody (BLR035F) [NBP2-76406] - Detection of human VISTA (red) in FFPE ovarian carcinoma by IHC-IF. Antibody: Rabbit anti-VISTA recombinant monoclonal [BLR035F] NBP2-76406. Secondary: HRP-conjugated goat anti-rabbit IgG. Substrate: Opal™. Counterstain: DAPI (blue).

Immunohistochemistry-Paraffin: VISTA/B7-H5/PD-1H Antibody (BLR035F) [NBP2-76406] - Detection of human VISTA in FFPE lung carcinoma by IHC. Antibody: Rabbit anti-VISTA recombinant monoclonal [BLR035F] (NBP2-76406). Secondary: HRP-conjugated goat anti-rabbit IgG. Substrate: DAB.

Flow Cytometry: VISTA/B7-H5/PD-1H Antibody (BLR035F) [NBP2-76406] - Rabbit anti-VISTA recombinant monoclonal [BLR035F] (right) or isotype control (left). Secondary: DyLight® 488-conjugated goat anti-rabbit IgG.

Immunocytochemistry/Immunofluorescence: VISTA/B7-H5/PD-1H Antibody (BLR035F) [NBP2-76406] - Detection of human VISTA in FFPE NCI-H226 cells by ICC. Antibody: Rabbit anti-VISTA recombinant monoclonal [BLR035F] (NBP2-76406). Secondary: HRP-conjugated goat anti-rabbit IgG. Substrate: DAB.

Immunoprecipitation: VISTA/B7-H5/PD-1H Antibody (BLR035F) [NBP2-76406] - Detection of human VISTA by WB of immunoprecipitates from Jurkat lysate. Antibodies: Rabbit anti-VISTA recombinant monoclonal [BLR035F] (NBP2-76406) and another rabbit anti-VISTA recombinant monoclonal.

• Reactivity: Hu
• Applications: WB, Flow, ICC/IF, IHC, IP, mIF
• Clone: BLR035F
• Clonality: Monoclonal
• Host: Rabbit
• Conjugate: Unconjugated

VISTA/B7-H5/PD-1H Antibody (BLR035F) Summary

• Additional Information: Recombinant Monoclonal Antibody.
• Immunogen: Immunogen sequence between amino acids 250 and 311 (C-Term)
• Isotype: IgG
• Clonality: Monoclonal
• Host: Rabbit
• Gene: VSIR
• Purity: Antigen Affinity-purified

Applications/Dilutions

• Dilutions:
  • Flow Cytometry Fixed in 4% formaldehyde and permeabilized with 90% methanol. 2 µl
  • Immunocytochemistry/ Immunofluorescence 1:100 - 1:500
  • Immunohistochemistry 1:100 - 1:500
  • Immunohistochemistry-Paraffin 1:100 - 1:500
  • Immunoprecipitation 20 ul/1mg lysate
  • Multiplex Immunofluorescence 1:250
  • Western Blot 1:1000
• Application Notes: Epitope retrieval with citrate buffer pH6.0 is recommended for FFPE tissue sections.

Packaging, Storage & Formulations

• Storage: Store at 2 - 8 C / 1 year from date of receipt
• Buffer: Borate Buffered Saline (BBS) pH 8.2 with 0.1% BSA
• Preservative: 0.09% Sodium Azide
• Purity: Antigen Affinity-purified

Notes

This Product sold under License from Bethyl Laboratories, Inc.

Alternate Names for VISTA/B7-H5/PD-1H Antibody (BLR035F)

• 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

Background

VISTA is an immunoregulatory receptor which inhibits the T-cell response (PubMed:24691993). It may promote differentiation of embryonic stem cells, by inhibiting BMP4 signaling (By similarity). VISTA may stimulate MMP14-mediated MMP2 activation

Limitations

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.

Bioinformatics

• Gene Symbol: VSIR