p70 S6 Kinase (T412E), active, Recombinant (p70 S6K)

A truncated, N-terminus His-tagged fusion protein corresponding to aa 1-421 of human p70 S6 Kinase (containing a T412E mutation and a D18H mutation) expressed in Sf21 cells. \nThe T412E mutant is more efficiently activated by PDK1 than is the wild type p70 S6 Kinase.\n\nSpecific Activity: ~200U/mg\n\nUnit Definition: \nOne unit of p70 S6 kinase activity is defined as 1nmol phosphate incorporated into 100uM S6 Kinase/Rsk-2 Substrate Peptide 2 (KKRNRTLTK) per minute at 30 degrees C with a final ATP concentration of 100uM. \n\nQuality Control Testing:\nMS Tryptic Fingerprint: Confirmed identity as p70 S6 Kinase with 32% amino acid coverage of the translated sequence.\n\nKinase Assay: 8.75-87.50ng phosphorylated 100uM S6 Kinase/Rsk-2 Substrate Peptide 2 in the assay described.\n\nStorage and Stability: \nMay be stored at -20 degrees C for short-term only. For long-term storage, store at -70 degrees C. Aliquots are stable for at least 6 months at -70 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.

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SPECIFICATIONS

Catalog Number

P1001-37B

Size

10ug

Form

Supplied as a liquid in 50mM Tris-HCl, pH 7.5, 270mM sucrose, 0.15M sodium chloride, 1mM benzamidine, 0.2mM PMSF, 0.1mM EGTA, 0.1% 2-mercaptoethanol, 0.03% Brij-35.

Purity

~ 90%. Purified by Ni-NTA agarose, activated with PDK1 and re-purified using Heparin Sepharose Chromatography. Purity is verified by SDS-PAGE.

References

1. Price, et al., J. Biol. Chem. 264: 13,825-13,833 (1989). 2. Alessi, D.R., et al., Current Biology 8: 69-81 (1997). 3. Thomas, G., Biochem. Soc. Trans. 20: 678-681 (1992). 4. Reinhard, C., et al., Proc. Natl. Acad. Sci. USA 85: 4052-4056 (1992). 5. Lehman, J.A., et al., J. Biol. Chem. 278: 28,130-28,138 (2003).