STAT (signal transducers and activators of transcription) can be selectively phosphorylated on tyrosine and serine residues. The STAT proteins are highly conserved in their amino termini, but have divergent carboxy termini, thought to be essential for their selective activity. STAT5 can be activated by a variety of different agents including prolactin, IL-2, IL-3, IL-7, IL-5, growth hormone and erythropoietin. The highly homologous STAT5A and STAT5B proteins are essential mediators of prolactin and growth hormone effects and have been implicated in physiological functions including cytokine control of apoptosis, growth, reproduction, lactation, immune function, differentiation and somatic growth.
Suitable for use in Western Blot and Immunoprecipitation. Other applications have not been tested.
Western Blot: 1:1000-1:2000 dilution detected STAT5B in RIPA lysates from Nb2-SP cells. Nb2-SP cell lysate (20ug) was resolved by electrophoresis, transferred to nitrocellulose and probed with S7972-15 (1:2000). Proteins were visualized using a goat anti-rabbit secondary antibody conjugated to HRP and a chemiluminescence detection system.
Immunoprecipitation: 4ul immunoprecipitated STAT5B from 500ug of Nb2-SP RIPA lysate.
Immunoprecipitation/Immunoblot Analysis: STAT5B was immunoprecipitated from Nb2-SP cell lysate (500 ug) using 4ul S7972-15. The reduced immuno-complex was resolved by electrophoresis, transferred to nitrocellulose and probed with S7972-15 (1:2000). Proteins were visualized using a goat anti-rabbit secondary antibody conjugated to HRP and a chemilum-inescence detection system.
Optimal dilutions to be determined by the researcher.
Storage and Stability:
May be stored at 4 degrees C for short-term only. For long-term storage and to avoid repeated freezing and thawing, aliquot and add glycerol (40-50%). Freeze at -20 degrees C. Aliquots are stable for at least 12 months at -20 degrees C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.