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Detection of Human Phospho-Paxillin (Y31) by Western Blot. Western blot shows lysates of A431 human epithelial carcinoma cell line, Jurkat human acute T cell leukemia cell line, and Daudi human Burkitt's lymphoma cell line untreated (-) or treated (+) with 1 mM Pervanadate (PV) for 30 minutes. PVDF membrane was probed with 0.1 μg/mL of Mouse Anti-Human Phospho-Paxillin (Y31) Antigen Affinity-purified Monoclonal Antibody (Catalog # MAB61641) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF007). Specific bands were detected for Phospho-Paxillin (Y31) at approximately 65 to 68 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Paxillin in HUVEC Human Cells. Paxillin phosphorylated at Y31 was detected in immersion fixed HUVEC human umbilical vein endothelial cells using Mouse Anti-Human Phospho-Paxillin (Y31) Antigen Affinity-purified Monoclonal Antibody (Catalog # MAB61641) at 10 μg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights? 557-conjugated Anti-Mouse IgG Secondary Antibody (red, upper panel; Catalog # NL007) and counterstained with DAPI (blue, lower panel). Specific staining was localized to nuclei and focal adhesions. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.