Human TRAILR1/TNFRSF10A Antibody

Detection of Human TRAIL R1/TNFRSF10A by Western Blot. Western blot shows lysates of TF-1 human erythroleukemic cell line. PVDF membrane was probed with 1 µg/mL of Goat Anti-Human TRAIL R1/TNFRSF10A Antigen Affinity-purified Polyclonal Antibody (Catalog # AF347) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF019). A specific band was detected for TRAIL R1/TNFRSF10A at approximately 50 kDa (as indicated). This experiment was conducted under reducing conditions.

TRAIL R1/TNFRSF10A Inhibition of TRAIL/TNFSF10-induced Cytotoxicity and Neutralization by Human TRAIL R1/TNFRSF10A Antibody. In the presence of a cross-linking antibody, Mouse polyHistidine Monoclonal Antibody (Catalog # MAB050) and the metabolic inhibitor actinomycin D (1 µg/mL), Recombinant Human TRAIL R1/TNFRSF10A Fc Chimera (Catalog # 347-DR) inhibits Recombinant Human TRAIL/TNFSF10 (Catalog # 375-TL) induced cytotoxicity in the L-929 mouse fibroblast cell line in a dose-dependent manner (orange line), as measured by crystal violet staining. Under these conditions, inhibition of Recombinant Human TRAIL/TNFSF10 (12 ng/mL) activity elicited by Recombinant Human TRAIL R1/TNFRSF10A Fc Chimera (10 ng/mL) is neutralized (green line) by increasing concentrations of Goat Anti-Human TRAIL R1/ TNFRSF10A Antigen Affinity-purified Polyclonal Antibody (Catalog # AF347). The ND₅₀ is typically 0.02-0.055 µg/mL.

TRAIL R1/TNFRSF10A in Human Brain. TRAIL R1/TNFRSF10A was detected in immersion fixed paraffin-embedded sections of human brain using Goat Anti-Human TRAIL R1/TNFRSF10A Antigen Affinity-purified Polyclonal Antibody (Catalog # AF347) at 10 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Goat HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS008) and counterstained with hematoxylin (blue). Specific staining was localized to cytoplasm in neurons. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.

Detection of Human TRAILR1/TNFRSF10A by Western Blot B3GNT2 disrupts ligand–receptor interactions between tumor and T cells.a Cell survival against T cell cytotoxicity (top) and T cell IFN gamma secretion (bottom) in A375 cells overexpressing B3GNT2 or GFP that have been treated with different concentrations of kifunensine. Kifunensine was used to pretreat A375 cells and was present during co-culture with T cells at E:T ratio of 3. Kifunensine-treated cells that were co-cultured with ESO T cells were compared to kifunensine-treated cells cultured in parallel without T cells to determine percent survival. N = 6. Two-tailed t tests were performed. b Tomato lectin IP of A375 cells overexpressing GFP or B3GNT2 followed by western blot for different B3GNT2 target proteins. 2% of the input and no lectin IP controls are shown. Data are representative of two independent experiments. c Western blots of A375 cells overexpressing B3GNT2 or GFP that were treated with kifunensine (KIF) or benzyl-2-acetamido-2-deoxy-alpha -D-galactopyranoside (BAG) to remove N- or O-glycosylation respectively. Data are representative of two independent experiments. d Histograms (top) and corresponding median fluorescence intensity (MFI; bottom) showing binding of recombinant T cell proteins to A375 cells measured by flow cytometry. A375 cells were overexpressing GFP or B3GNT2 and treated with KIF or BAG. N = 3. Two-tailed t tests were performed. e Schematic showing the tumor cell surface ligands and receptors modified by B3GNT2 to disrupt interactions with T cells that mediate cytotoxicity. All values are mean ± s.e.m. ns not significant. Source data are provided in Source Data 4. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/35338135), licensed under a CC-BY license. Not internally tested by R&D Systems.