Human Nectin-2/CD112 Antibody

Detection of Human Nectin‑2/CD112 by Western Blot. Western blot shows lysates of 293T human embryonic kidney cell line, A549 human lung carcinoma cell line, Daudi human Burkitt's lymphoma cell line, and Raji human Burkitt's lymphoma cell line. PVDF membrane was probed with 0.1 µg/mL of Goat Anti-Human Nectin-2/CD112 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2229) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). A specific band was detected for Nectin-2/CD112 at approximately 60-75 kDa (as indicated). Daudi human Burkitt's lymphoma cell line and Raji human Burkitt's lymphoma cell line are shown as negative controls. GAPDH (Catalog # AF5718) is shown as a loading control.This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Nectin‑2/CD112 in MCF‑7 Human Cell Line. Nectin-2/CD112 was detected in immersion fixed MCF-7 human breast cancer cell line using Goat Anti-Human Nectin-2/CD112 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2229) at 1.7 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red; Catalog # NL001) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

Nectin‑2/CD112 in Human Liver. Nectin-2/CD112 was detected in immersion fixed paraffin-embedded sections of human liver using Goat Anti-Human Nectin-2/CD112 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2229) at 3 µg/mL for 1 hour at room temperature followed by incubation with the Anti-Goat IgG VisUCyte™ HRP Polymer Antibody (Catalog # VC004). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to cell membrane. View our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.

Detection of Human Nectin‑2/CD112 by Simple Western^TM. Simple Western lane view shows lysates of HEK293T human embryonic kidney cell line, A549 human lung carcinoma cell line, Daudi human Burkitt's lymphoma cell line, and Raji human Burkitt's lymphoma cell line, loaded at 0.2 mg/mL. A specific band was detected for Nectin-2/CD112 at approximately 71-86 kDa (as indicated) using 20 µg/mL of Goat Anti-Human Nectin-2/CD112 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2229) followed by 1:50 dilution of HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF109). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.

Detection of Human Nectin-2/CD112 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 publication (https://pubmed.ncbi.nlm.nih.gov/35338135), licensed under a CC-BY license. Not internally tested by R&D Systems.