Recombinant Human RPS6KB1 p70S6K His Active *di

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Hersteller Cell Sciences
Kategorie
Typ Proteins
Specific against Human
Menge 10 ug
Host Insect
ArtNr CS-CRP104B
eClass 6.1 34160400
eClass 9.0 42020190
Lieferbar
Description
Recombinant full-length human p70S6K containing N-terminal His tag was expressed by baculovirus in Sf 9 insect cells. Activation of cell growth leads to the multiple phosphorylation of 40S ribosomal protein S6. The kinase responsible for controlling this event is termed p70S6K. Northern blot analysis shows p70S6K to be ubiquitously expressed in human adult tissues. p70S6K is activated by serum stimulation, and the serum-induced activation is inhibited by wortmannin and rapamycin. p70S6k activity undergoes changes in the cell cycle and increases 20-fold in G1 cells released from G0. The kinase is reactivated 10-fold when cells released from a nocodazole-induced metaphase block enter G1 of the next cell cycle. The immunosuppressive agent rapamycin induces inactivation of p70s6k with no effect on other mitogen-activated kinases. The principal target of rapamycin-induced p70s6k inactivation is T389 site on p70S6K, which is located in an unusual hydrophobic sequence outside the catalytic domain. Mutation of T389 to alanine ablates kinase activity, whereas mutation to glutamic acid confers constitutive kinase activity and rapamycin resistance. p70S6K activation by growth factor requires phosphorylation by various inputs on multiple sites. p70S6K activation requires sequential phosphorylations at proline-directed residues in the putative autoinhibitory pseudosubstrate domain, as well as threonine 389. Threonine 229, a site in the catalytic loop is phosphorylated by PDK-1. Activation of p70S6K requires a phosphoinositide 3-kinase (PI3-K)-dependent signal(s).
Formulation
Recombinant protein in storage buffer (50 mM Tris-HCl + 150 mM NaCl + 0.25 mM DTT + 0.1 mM EGTA + 0.1 mM EDTA + 0.1 mM PMSF + 25% glycerol, pH, 7.5).
Storage and Stability
Store product frozen at or below -70C. Stable for 1 year at -70C as undiluted stock. Aliquot to avoid repeated thawing and freezing.
Concentration
0.1 mg/ml
Source
Sf9 insect cells
Purity
1 ug of p70S6K protein was subjected to SDS-PAGE and Coomassie blue staining. The scan of the gel showed >90% purity of the p70S6K product, and the band was at ca.76 kDa (Fig. 2).
Specific Activity
80 nmol/min/mg: 80 nmol phosphate incorporated into S6K substrate peptide per minute per mg protein at 30C for 15 minutes using a final concentration of 50 uM ATP (0.83 uCi/assay). See QA/QC section for details.

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Menge: 10 ug
Lieferbar: In stock
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