The Kinase Enzyme Systems include a recombinant kinase enzyme, a substrate appropriate for the enzyme, a reaction buffer and supplemental reagents as needed. The p70S6K Kinase Enzyme System contains:
- p70S6K Kinase, 10μg (Human, recombinant full-length). MW: ~76kDa.
- S6K Substrate (KRRRLASLR); derived from human 40S ribosomal protein S6 (amino acid 230–238).
- Reaction Buffer, DTT.
Recombinant full-length human p70S6K was expressed by baculovirus in Sf9 insect cells using an N-terminal His tag. 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 during 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).
p70S6K NCBI Database Entry.
The p70S6K Kinase Enzyme System can be purchased with or without the ADP-Glo™ Kinase Assay reagents. Used together, the ADP-Glo™ Kinase Assay + Kinase Enzyme Systems provide a convenient method for profiling the effect of lead compounds on kinase activity. Assay advantages include broad dynamic range, ease of use and high sensitivity. Kinase Enzyme Systems are manufactured by SignalChem. Bulk quantities available upon request.
Use with ADP-Glo™ Kinase Assay
The ADP-Glo™ Kinase Assay is a luminescent kinase assay that measures ADP formed from a kinase reaction; ADP is converted into ATP, which is a substrate in a reaction catalyzed by Ultra-Glo™ Luciferase that produces light. The luminescent signal positively correlates with ADP amount and kinase activity. The assay is well suited for measuring the effects of chemical compounds on the activity of a broad range of purified kinases, making it ideal for both primary screening as well as kinase selectivity profiling. The ADP-Glo™ Kinase Assay can be used to monitor the activity of virtually any ADP-generating enzyme (e.g., kinase or ATPase) using up to 1mM ATP.
See all Kinase Enzyme Systems available from Promega.