Recombinant Anti-VISTA antibody [EPR21050] (ab230950)

Immunohistochemical analysis of paraffin-embedded human lung cancer tissue labeling VISTA with ab230950 at 1/1000 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP) secondary antibody. Membranous and cytoplasmic staining in tumor-infiltrating leukocytes of human lung cancer (PMID: 28236118, PMID: 28507801) is observed. Counterstained with hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).

Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

Blocking/Dilution buffer: 5% NFDM/TBST.

Exposure times: Lanes 1-2: 32 seconds; Lanes 3-4: 6 seconds.

The 50-60 kda band observed is the glycosylated form of VISTA, consistent with what has been described in the literature (PMID: 29216931).

Immunohistochemical analysis of paraffin-embedded human spleen tissue labeling VISTA with ab230950 at 1/1000 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP) secondary antibody. Membranous and cytoplasmic staining in leukocytes of human spleen (PMID: 21383057, PMID: 24691993) is observed. Counterstained with hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).

Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

VISTA was immunoprecipitated from 0.35 mg DU145 (Human prostate carcinoma epithelial cell) whole cell lysate with ab230950 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab230950 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/5000 dilution.
Lane 1: DU154 whole cell lysate 10 µg (Input).
Lane 2: ab230950 IP in DU154 whole cell lysate.
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab230950 in DU154 whole cell lysate.
Blocking/Dilution buffer: 5% NFDM/TBST.
Exposure time: 3 minutes.

Tissue Microarrays stained for "Anti-VISTA antibody [EPR21050]” using "ab230950" in immunohistochemical analysis. This table provides a detailed overview of positive (tick mark) and negative (cross mark) staining per sample type tested. The sections were pre-treated using Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0). The sections were incubated with ab230950 at +4°C overnight followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP polymer).