Human Phospho-PYK2/FAK2 (Y402) Antibody

Detection of HumanPhospho-PYK2/FAK2 (Y402) by Western Blot.

Western blot shows lysates of Raji human Burkitt's lymphoma cell line and Jurkat human acute T cell leukemia cell line untreated (-) or treated (+) with 1 mM Pervanadate (PV) for 30 minutes and 10 µg/mL Mouse Anti-Human CD3e Monoclonal Antibody (Catalog # MAB100) for 15 minutes. PVDF membrane was probed with 0.5 µg/mL of Mouse Anti-Human Phospho-PYK2/FAK2 (Y402) Monoclonal Antibody (Catalog # MAB6210) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF018). Specific bands were detected for Phospho-PYK2/FAK2 (Y402) at approximately 105-115 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Detection of Human Phospho-PYK2/FAK2 (Y402) by Simple Western^TM.

Simple Western lane view shows lysates of Raji human Burkitt's lymphoma cell line untreated (-) or treated (+) with 0.2 mg/mL Pervanadate (PV) for 30 minutes, loaded at 0.2 mg/mL. A specific band was detected for Phospho-PYK2/FAK2 (Y402) at approximately 113 kDa (as indicated) using 5 µg/mL of Mouse Anti-Human Phospho-PYK2/FAK2 (Y402) Monoclonal Antibody (Catalog # MAB6210). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.

Detection of Phospho-PYK2/FAK2 (Y402) by Western Blot

Galectin-3 expression induces activation of PYK2, STAT1 and GSK3 alpha/ beta signalling. Expression of 37 protein kinases in SW620 cells in response to 10 µg/ml galectin-3 or BSA for 0.5 h was assessed by Proteome Profiler Human Phospho-Kinase Array (A, Percentage changes of the kinases in cell response to galectin-3 in comparison to control are shown at the bottom panel). The presence of galectin-3 increases the phosphorylation of PYK2, GSK3 alpha/ beta, and STAT1 and decreases phosphorylation of STAT3. SW620 cells treated with 10 µg/ml galectin-3 for different times were assessed by immunoblotting using antibodies against p-PYK2, p-STAT-1, p-GSK3 alpha/ beta or p-STAT-3 (B). The blots were striped and reprobed with antibodies against PYK2, STAT-1, GSK3 alpha/ beta or STAT-3. The band density was quantified and expressed as percentages of phospho-/non-phosphorylated proteins (C). In D and E, SW620 cells were treated with 10 µg/ml galectin-3 or BSA followed by introduction of GSK3 alpha/ beta inhibitor SB 216763 (SB) or PKY2 inhibitor PF-431396 (PF) for 15 min and the levels of phosphorylated PYK2, STAT-1, GSK3 alpha/ beta or STAT-3 were analysed by immunoblotting. The blots were striped and reprobed with antibodies against PYK2, STAT-1, GSK3 alpha/ beta or STAT-3. The densities of the blots from three independent experiments were quantified and are expressed as the percentage of phosphorylated/non-phosphorylated levels of each protein. ***P < 0.001, **P < 0.01, *P < 0.05 (ANOVA). Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/37055381), licensed under a CC-BY license. Not internally tested by R&D Systems.