Human N-Cadherin Antibody

Detection of Human, Mouse, and Rat N‑Cadherin by Western Blot. Western blot shows lysates of A549 human lung carcinoma cell line, C2C12 mouse myoblast cell line, and rat brain tissue. PVDF membrane was probed with 1 µg/mL of Mouse Anti-Human N-Cadherin Monoclonal Antibody (Catalog # MAB13881) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF007). A specific band was detected for N-Cadherin at approximately 130 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Detection of N-Cadherin in HeLa Human Cell Line by Flow Cytometry. HeLa human cervical epithelial carcinoma cell line was stained with Mouse Anti-Human N-Cadherin Monoclonal Antibody (Catalog # MAB13881, filled histogram) or isotype control antibody mouse IgM (open histogram), followed by Phycoerythrin-conjugated Anti-Mouse IgM Secondary Antibody (Catalog # F0116).

N‑Cadherin in Human Brain. N-Cadherin was detected in immersion fixed paraffin-embedded sections of Alzheimer's human brain using Mouse Anti-Human N-Cadherin Monoclonal Antibody (Catalog # MAB13881) at 15 µg/mL overnight at 4 °C. Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using Antigen Retrieval Reagent-Basic (Catalog # CTS013). Tissue was stained using the Anti-Mouse HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS002) and counterstained with hematoxylin (blue). Specific staining was localized to cytoplasm of neurons. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.

Detection of N-Cadherin by Western Blot DCN inhibits E-cadherin expression and EGFR pathway activation in IBC.a DCN suppresses E-cadherin expression and EGFR signaling in IBC cells. Expression levels of E-cadherin and EGFR are decreased in DCN-overexpressing MDA-IBC3, SUM190, SUM149, and BCX010 cells; also, the phosphorylation of EGFR (pEGFR) and ERK1/2 (pERK1/2) was suppressed in DCN-overexpressing IBC cell lines. Total ERK1/2 (tERK1/2) remains unchanged. GAPDH served as a loading control. b Treatment of IBC cells with DCN protein (4 or 8 μg/mL) for 2 h suppresses E-cadherin expression and EGFR pathway activation. Tubulin served as a loading control. c and d Western blot validation of E-cadherin and EGFR downregulation in tumor samples obtained from mammary fatpad transplantation of control or DCN-overexpressing MDA-IBC3 (c) or SUM149 (d) cells. e and f Immunohistochemical staining validation of E-cadherin and EGFR downregulation in tumor samples obtained from mammary fatpad transplantation of control or DCN-overexpressing MDA-IBC3 (e) or SUM149 (f) cells. Scale bar: 100 μm. g DCN inhibits EGFR signaling in IBC cells independently of EGF stimulation. DCN-overexpressing and control IBC cell lines were stimulated with 50 ng/mL EGF for the indicated number of hours, and total cell lysates were analyzed by western blotting. Both the total levels and the phosphorylation levels of EGFR and ERK1/2 were detected by western blotting. Tubulin served as a loading control. h DCN-mediated inhibition of E-cadherin does not affect expression of epithelial–mesenchymal transition markers. Cell lysates containing 40 μg of total protein were analyzed by western blotting with anti-E-cadherin, fibronectin, vimentin, and DCN antibodies. GAPDH served as a loading control. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/33452400), licensed under a CC-BY license. Not internally tested by R&D Systems.