p70S6K Kinase Enzyme System

The Kinase Enzyme Systems include a recombinant kinase enzyme, a substrate appropriate for the enzyme, a reaction buffer and supplemental reagents as needed. The PDK1 Kinase Enzyme System contains:

• PDK1 Kinase, 10μg (Human, recombinant full-length). MW: ~67kDa.
• PDKtide (KTFCGTPEYLAPEVRREPRILSEEEQEMFRDFDYIADWC) Substrate; derived from two human proteins: residues 114 are based on AKT1 (307320) and residues 1639 are based on PKN2/PRK2 (961984).
• Reaction Buffer, DTT.

Recombinant full-length human PDK1 was expressed by baculovirus in Sf9 insect cells using an N-terminal His tag. PDK1 (3-phosphoinositide-dependent protein kinase) is activated by the presence of PtdIns(3,4,5)P3 or PtdIns(3,4)P2. PDK1 then activates protein kinase B (PKB), which in turn, inactivates glycogen synthase kinase-3 (GSK3). The phosphorylation of other proteins by PKB and GSK3 is likely to mediate many of the intracellular actions of insulin. Thus, PDK1 plays a key role in mediating many of the actions of the second messenger(s) PtdIns(3,4,5)P3 and/or PtdIns(3,4)P2. The human PDK1 is a 556-residue monomeric enzyme comprising of a catalytic domain that is most similar to the PKA, PKB and PKC subfamily of protein kinases.

PDK1 NCBI Database Entry.

The PDK1 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.