Human Phospho-STAT1 (Y701) Antibody

Detection of Human Phospho-STAT1 (Y701) by Western Blot.

Western blot shows lysates of Daudi human Burkitt's lymphoma cell line untreated (-) or treated (+) with Recombinant Human IFN-aA (Catalog # 11100-1) for 20 minutes. PVDF membrane was probed with 0.25 µg/mL of Rabbit Anti-Human Phospho-STAT1 (Y701) Monoclonal Antibody (Catalog # MAB2894) followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # HAF008). A specific band was detected for Phospho-STAT1 (Y701) at approximately 90 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Detection of Phospho-STAT1 (Y701) in IFN alpha-treated Daudi Human Cell Line by Flow Cytometry.

Daudi human Burkitt's lymphoma cell line was unstimulated (open histogram) or treated with 500 U/mL rhIFN-alpha for 20 minutes (filled histogram) and stained with Rabbit anti-Human Phospho-STAT1 (Y701) Molyclonal Antibody (Catalog # MAB2894) followed by APC-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # F0111). To facilitate intracellular staining, cells were fixed with paraformaldehyde and permeabilized with methanol. View our protocol for Staining Intracellular Molecules.

Detection of Phospho-STAT1 (Y701) 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.