Anti-phospho-STAT5a,b (Ser726/Ser731) (Signal Transducers and Activators of Transcription 5a,b)

Many hormones and cytokines selectively activate transcription factors of the STAT gene family. STAT5 was originally cloned as a prolactin-stimulated DNA-binding protein, but is also activated by non-lactogenic cytokines in many cell types. Phosphorylation of a conserved tyrosine residue is required for STAT dimerization, nuclear translocation, and specific DNA binding. Serine residues located in a conserved PSP motif of STAT5A (S726) and STAT5B (S731) are also major phosphorylation sites. Interestingly, control of the phosphorylation of the PSP motif differs between the STAT5A (S725) and STAT5B (S730) serine sites. Applications: Suitable for use in Immunoprecipitation/Western Blot. Other applications have not been tested. Recommended Dilutions: Immunoprecipitation/Western Blot Analysis: 1:500-1:2000 detected phosphorylated STAT5A following immunoprecipitation of STAT5A from RIPA lysates of Nb2-Sp cells that had been treated with prolactin, 4ul of S7972-10 was used to immunoprecipitate total STAT5A. Proteins were visualized using an IgG (HRP) goat anti-rabbit secondary antibody and a chemiluminescence detection system. Phosphospecificity was confirmed by dephosphorylation of the STAT5a protein with l-phosphatase., Optimal dilutions to be determined by the researcher. Recommended Secondary Antibodies: I1904-39: IgG, X-Adsorbed (HRP) Pab Gt xRb, I1904-40A: IgG, H&L, X-Adsorbed (HRP) Pab Gt xRb, I1904-46Q: IgG, H&L (HRP) Pab Gt xRb, Storage and Stability: May be stored at 4°C for short-term only. Aliquot to avoid repeated freezing and thawing. Store at -20°C. Aliquots are stable for 12 months after receipt. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.