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7577
HTScan® PDK1 Kinase Assay Kit

HTScan® PDK1 Kinase Assay Kit #7577

This product is discontinued

Kinase Assay - Radiometric Image 1

Figure 1. 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.

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Kinase Assay - DELFIA Image 2

Figure 3. Dose dependence curve of PDK1 kinase activity: DELFIA® data generated using Phospho-PKA C (Thr197) Antibody #4781 to detect phosphorylation of substrate peptide (#1024) by PDK1 kinase. In a 50 µl reaction, increasing amounts of PDK1 and 3.0 µM substrate peptide were used per reaction at room temperature for 30 minutes. (DELFIA® is a registered trademark of PerkinElmer, Inc.)

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Kinase Assay - DELFIA Image 3

Figure 5. Staurosporine inhibition of PDK1 kinase activity: DELFIA® data generated using Phospho-PKA C (Thr197) Antibody #4781 to detect phosphorylation of PDK1 substrate peptide (#1024) by PDK1 kinase. In a 50 µl reaction, 50 ng PDK1, 3.0 µM substrate peptide, 20 µM ATP and increasing amounts of staurosporine were used per reaction at room temperature for 30 minutes. (DELFIA® is a registered trademark of PerkinElmer, Inc.)

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Kinase Assay - DELFIA Image 4

Figure 4. Peptide concentration dependence of PDK1 kinase activity: DELFIA® data generated using Phospho-PKA C (Thr197) Antibody #4781 to detect phosphorylation of substrate peptide (#1024) by PDK1 kinase. In a 50 µl reaction, 50 ng of PDK1 and increasing concentrations of substrate peptide were used per reaction at room temperature for 30 minutes. (DELFIA® is a registered trademark of PerkinElmer, Inc.)

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Kinase Assay - DELFIA Image 5

Figure 2. Time course of PDK1 kinase activity: DELFIA® data generated using Phospho-PKA C (Thr197) Antibody #4781 to detect phosphorylation of PDK1 substrate peptide (#1024) by PDK1 kinase. In a 50 µl reaction, 50 ng PDK1 and 3.0 µM substrate peptide were used per reaction. (DELFIA® is a registered trademark of PerkinElmer, Inc.)

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Product Includes Quantity (with Count) Solution Color
Kinase Buffer (10X) 9802 1 x 15 ml
ATP (10 mM) 9804 1 x 1 ml

The kit provides a means of performing kinase activity assays with recombinant human PDK1 kinase. It includes active PDK1 kinase (supplied as a GST fusion protein), a biotinylated peptide substrate and a phospho-serine/threonine antibody for detection of the phosphorylated form of the substrate peptide.

Molecular Formula:

Biotin-peptide: 2,205 Daltons. PDK1 Kinase: 67 kDa.

Peptide Core Sequence:

RTWT*LCG

Phosphoinositide-dependent protein kinase 1 (PDK1) plays a central role in many signal transduction pathways (1,2) including the activation of 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.

  1. Belham, C. et al. (1999) Curr. Biol. 9, R93-R96.
  2. Toker, A. and Newton, A.C. (2000) Cell 103, 185-188.
  3. Williams, M.R. et al. (2000) Curr. Biol. 10, 439-448.
Entrez-Gene Id
5170
Swiss-Prot Acc.
O15530
For Research Use Only. Not For Use In Diagnostic Procedures.

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
DELFIA is a registered trademark of PerkinElmer, Inc.

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