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Encompassing Amino Acids: 1-421
Applications: Useful for the study of enzyme kinetics, screening inhibitors, and selectivity profiling.
Assay Conditions: Enzyme reaction is conducted in a buffer containing 50 mM HEPES (pH 7.5), 10 mM MgCl2, 1 mM EGTA, 200 µM ATP, 0.01% Brij-35 and 2 µM substrate (SER/THR 7, Invitrogen) at room temperature for 1 hour.
Background: p70S6K is responsible for the phosphorylation of 40S ribosomal protein S6 and is ubiquitously expressed in human adult tissues. p70S6K is activated by serum stimulation and this 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. p70S6K activation requires sequential phosphorylations at proline-directed residues in the putative autoinhibitory pseudosubstrate domain, as well as threonine 389 a site phosphorylated by phosphoinositide-dependent kinase 1 (PDK-1).
Description: Human p70S6Kb1, also known as ribosomal protein S6 kinase, 70 kDa, polypeptide 1 (RPS6KB1), GenBank Accession No. NM_003161, amino acid 1-421 with N-terminal GST tag, MW=74 kDa, expressed in a Baculovirus infected Sf9 cell expression system.
Format: Aqueous buffer solution
Formulation: 25 mM Tris-HCl, pH 8.0, 100 mM NaCl, 0.05% Tween-20, 30% glycerol, and 3 mM DTT.
Genbank: NM_003161
Host Cell Line: Sf9 cells
Storage Stability: At least 6 months at -80°C.
Tags: N-terminal GST-tag
Uniprot: P23443
Warnings: Avoid freeze/thaw cycles.
Biosafety Level: Not applicable (BSL-1)
References: 1. Di Paolo S. et al., Am. J. Kidney Dis. 49 (3), 462-470 (2007).
2. Fang C.X. et al., J. Exp. Neurol. 203 (1), 196-204 (2007).
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