Product Pathways - Akt Signaling
PDK1 Kinase #7386
Cell Signaling Technology offers a full line of protein kinases, substrates, antibody detection reagents and HTScan® kits. Browse our "Reagents for High-Throughput Screening" product listing or contact us at drugdiscovery@cellsignal.com.
Description
Purified recombinant full length human PDK1 kinase.
Source / Purification
The kinase protein was produced using a baculovirus expression system with a construct expressing full length human PDK1 (Met1-Gln556) (GenBank Accession No. NM_002613) with an amino-terminal His tag. The protein was purified by Immobilized Metal Affinity Chromatography (IMAC).
Gel Staining
Figure 1. The purity of the PDK1 fusion protein was analyzed using SDS/PAGE followed by Coomassie stain.
Kinase Assay - Radiometric
Figure 2. PDK1 kinase activity was measured in a radiometric assay using the following reaction conditions: 5 mM MOPS, pH 7.2, 2.5 mM β-glycerophosphate, 1 mM EGTA, 0.4 mM EDTA, 5 mM MgCl2, 0.05 mM DTT, 50 ng/μl BSA, 50 μM ATP, Substrate: PDKtide 400 ng/μL, and recombinant PDK1: variable.
Quality Control
The theoretical molecular weight of the PDK1 protein is 67 kDa. The purified kinase was quality controlled for purity using SDS-PAGE followed by Coomassie stain [Fig.1]. PDK1 kinase activities were determined using a radiometric assay [Fig.2].
Background
Phosphoinositide-dependent protein kinase 1 (PDK1) plays a central role in many signal transduction pathways (1,2), activating Akt and the PKC isoenzymes p70 S6 kinase and RSK (3). Through its effects on these kinases, PDK1 is involved in the regulation of a wide variety of processes, including cell proliferation, differentiation and apoptosis.
- Belham, C. et al. (1999) Curr. Biol. 9, R93-R96.
- Toker, A. and Newton, A.C. (2000) Cell 103, 185-188.
- Williams, M.R. et al. (2000) Curr. Biol. 10, 439-448.
Application References
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Companion Products
- 7577 HTScan® PDK1 Kinase Assay Kit
- 1024 PKA (Thr197) Biotinylated Peptide
- 4781 Phospho-PKA C (Thr197) Antibody
- 9802 Kinase Buffer (10X)
- 9804 ATP (10 mM)
- 9953 Staurosporine
This product is for in vitro research use only and is not intended for use in humans or animals. This product is not intended for use as therapeutic or in diagnostic procedures.
