STAT 5a,b, phosphorylated (Ser726/Ser731) (Signal Transducer and Activator 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.\n\nApplications: \nSuitable for use in Western Blot and Immunoprecipitation. Other applications have not been tested.\n\nRecommended Dilutions:\nImmunoprecipitation/Western Blot Analysis: 1:500-1:2000 dilution 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.\nOptimal dilutions to be determined by the researcher.\n\nRecommended Secondary Antibodies:\nI1904-39: IgG, X-Adsorbed (HRP) Pab Gt xRb\nI1904-40A: IgG, H&L, X-Adsorbed (HRP) Pab Gt xRb\nI1904-46Q: IgG, H&L (HRP) Pab Gt xRb\n\nStorage and Stability:\nMay be stored at 4 degrees C for short-term only. Aliquot to avoid repeated freezing and thawing. Store at -20 degrees C. Aliquots are stable for 12 months. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.

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SPECIFICATIONS

Catalog Number

S7972-12

Size

200ul

Applications

IP, WB

Hosts

Rabbit

Reactivities

Hum, Mouse, Rat

Form

Supplied as a liquid, 0.05% sodium azide, 30% glycerol.

P Type

Pab

Purity

Serum

Isotype

IgG

References

Yamashita H., et al., J. Biol. Chem. 273: 30218-30224, 1998. Kirken R.A., et al., J. Biol. Chem. 272: 14098-14103, 1997. Kirken R.A., et al., J. Biol. Chem. 272: 15459-15465, 1997.

Additional Info

Recognizes human phosphorylated STAT5A (S726) and STAT5B (S731) at ~95kD. Does not recognize STAT5B containing a S731A mutation in lysates of transfected Cos-1 cells, stimulated with prolactin. Species Crossreactivity: mouse and rat.