Human Fibronectin Antibody

Detection of Fibronectin in Human Breast Tumor via seqIF™ staining on COMET™​ Fibronectin was detected in immersion fixed paraffin-embedded sections of human Breast Tumor using Mouse Anti-Human Fibronectin Monoclonal Antibody (Catalog # MAB19182) at 15ug/mL 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™ 555 Goat anti-Mouse IgG Secondary Antibody at 1:100 at 37 ° Celsius for 2 minutes. (Yellow; Lunaphore Catalog # DR555MS) and counterstained with DAPI (blue; Lunaphore Catalog # DR100). Specific staining was localized to the cytoplasm. Protocol available in COMET™ Panel Builder.​​

Detection of Human Fibronectin by Western Blot. Western blot shows lysates of HepG2 human hepatocellular carcinoma cell line. PVDF membrane was probed with 2 µg/mL of Mouse Anti-Human Fibronectin Monoclonal Antibody (Catalog # MAB19182) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (HAF018). A specific band was detected for Fibronectin at approximately 300 kDa (as indicated). This experiment was conducted under reducing conditions and using Western Blot Buffer Group 1.

Fibronectin in HepG2 Human Cell Line. Fibronectin was detected in immersion fixed HepG2 human hepatocellular carcinoma cell line treated with monensin using Mouse Anti-Human Fibronectin Monoclonal Antibody (Catalog # MAB19182) at 3 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (red; Catalog # NL007) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm (punctate). View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

Fibronectin in Human Liver Cancer Tissue. Fibronectin was detected in immersion fixed paraffin-embedded sections of human liver cancer tissue using Mouse Anti-Human Fibronectin Monoclonal Antibody (Catalog # MAB19182) at 5 µg/mL for 1 hour at room temperature followed by incubation with the Anti-Mouse IgG VisUCyte™ HRP Polymer Antibody (Catalog # VC001). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to cytoplasm and plasma membrane in tumor cells. View our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.

Detection of Fibronectin by Western Blot KPA treatment decreases activation of human hepatic stellate LX-2 cells. Representative Western blots showing expression of endogenous (A) KISS1 protein (n = 5 biological replicates) and (B) KISS1R protein expression (n = 4 biological replicates). MDA-MB-231, SKBR3, and KISS1R-SKBR3 were used as reference for expression (n = 4). (C) Secreted kisspeptin protein in culture media measured by ELISA (N-4 biological replicates). (D–G) Changes in fibrogenic gene expression in response to KPA (3 nM, 48 h) +/− TGFb (5 ng/mL, 48 h). (n = 4–6 biological replicates) (H) Western blot analysis and (I,J) densitometric analyses of blots, showing changes in fibrogenic protein in response to KPA (3 nM, 72 h) +/− TGFb (5 ng/mL, 72 h). (n = 4 biological replicates) Results are expressed as mean +/− S.E.M. * p < 0.05 vs. control. ** p < 0.01 vs. control. *** p < 0.001 vs. control. **** p < 0.0001 vs. control. One-way ANOVA was conducted, followed by multiple comparison test. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/39404414), licensed under a CC-BY license. Not internally tested by R&D Systems.