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7621
HTScan® SGK3 Kinase Assay Kit

HTScan® SGK3 Kinase Assay Kit #7621

This product is discontinued

Kinase Assay - Radiometric Image 1

Figure 1. SGK3 kinase activity was measured in a radioisotopic filter binding assay using the following reaction conditions: 60 mM HEPES-NaOH, pH 7.5, 3 mM MgCl2, 3 mM MnCl2, 3 µM Na-orthovanadate, 1.2 mM DTT, 1 µM ATP, 2.5 µg/50 µl PEG20,000, Substrate: GST-GSK3 (14-27), variable and recombinant SGK3, variable.

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Kinase Assay - DELFIA Image 2

Figure 3. Dose dependence curve of SGK3 kinase activity: DELFIA® data generated using Phospho-PLK (Ser137) Antibody #5070 to detect phosphorylation of substrate peptide (#1300) by SGK3 kinase. In a 50 µl reaction, increasing amounts of SGK3 and 1.5 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 SGK3 kinase activity: DELFIA® data generated using Phospho-PLK (Ser137) Antibody #5070 to detect phosphorylation of SGK3 substrate peptide (#1300) by SGK3 kinase. In a 50 µl reaction, 50 ng SGK3, 1.5 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 SGK3 kinase activity: DELFIA® data generated using Phospho-PLK (Ser137) Antibody #5070 to detect phosphorylation of substrate peptide (#1300) by SGK3 kinase. In a 50 µl reaction, 50 ng of SGK3 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 SGK3 kinase activity: DELFIA® data generated using Phospho-PLK (Ser137) Antibody #5070 to detect phosphorylation of SGK3 substrate peptide (#1300) by SGK3 kinase. In a 50 µl reaction, 50 ng SGK3 and 1.5 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
Phospho-PLK (Ser137) Antibody 1 x 30 µl
ATP (10 mM) 9804 1 x 1 ml

The kit provides a means of performing kinase activity assays with recombinant full-length human SGK3 kinase. It includes active SGK3 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:

Peptide substrate, Biotin-PLK (Ser137): 1,945 Daltons. GST-SGK3 Kinase: 83 kDa.

Peptide Core Sequence:

RRS*LL

Serum and glucocorticoid-inducible kinase (SGK) is a serine/threonine kinase closely related to Akt (1). SGK is rapidly induced in response to a variety of stimuli, including serum, glucocorticoid, follicle stimulating hormone, osmotic shock, and mineralocorticoids. SGK activation can be accomplished via HGF PI3K-dependent pathways and by integrin-mediated PI3K-independent pathways (2,3). Induction and activation of SGK has been implicated in activating the modulation of anti-apoptotic and cell cycle regulation (4-6). SGK also plays an important role in activating certain potassium, sodium, and chloride channels, suggesting its involvement in the regulation of processes such as cell survival, neuronal excitability, and renal sodium excretion (2). SGK is negatively regulated by ubiquitination and proteasome degradation (7).

  1. Webster, M.K. et al. (1993) Mol Cell Biol 13, 2031-40.
  2. Kobayashi, T. and Cohen, P. (1999) Biochem J 339 ( Pt 2), 319-28.
  3. Park, J. et al. (1999) EMBO J 18, 3024-33.
  4. Brunet, A. et al. (2001) Mol Cell Biol 21, 952-65.
  5. Mikosz, C.A. et al. (2001) J Biol Chem 276, 16649-54.
  6. Hayashi, M. et al. (2001) J Biol Chem 276, 8631-4.
  7. Brickley, D.R. et al. (2002) J Biol Chem 277, 43064-70.
Entrez-Gene Id
23678
Swiss-Prot Acc.
Q96BR1
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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