Revision 1

#7621Store at -80C

Cell Signaling Technology

Orders: 877-616-CELL (2355) [email protected]

Support: 877-678-TECH (8324)

Web: [email protected] cellsignal.com

3 Trask LaneDanversMassachusetts01923USA
For Research Use Only. Not for Use in Diagnostic Procedures.
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UniProt ID:

#Q96BR1

Entrez-Gene Id:

23678

Product Information

Storage

Antibodies are supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA and 50% glycerol. Do not aliquot the antibodies. Peptides are supplied at 6 µM in 0.001% DMSO. Enzymes are supplied in 50 mM Tris-HCL (pH 8.0), 100 mM NaCl,
5 mM DTT, 15 mM reduced glutathione and 20% glycerol. Store at -80°C.
Keep enzymes on ice during use.
Avoid repeated freeze-thaw cycles.

Source / Purification

The GST-Kinase fusion protein was produced using a baculovirus expression system with a construct expressing full-length human SGK3 (Met1-Leu496) (GenBank Accession No. NM_013257.3) with an amino-terminal GST tag. The protein was purified by one-step affinity chromatography using glutathione-agarose.

Product Description

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.

Background

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.

Species Reactivity

Species reactivity is determined by testing in at least one approved application (e.g., western blot).

Cross-Reactivity Key

H: human M: mouse R: rat Hm: hamster Mk: monkey Vir: virus Mi: mink C: chicken Dm: D. melanogaster X: Xenopus Z: zebrafish B: bovine Dg: dog Pg: pig Sc: S. cerevisiae Ce: C. elegans Hr: horse GP: Guinea Pig Rab: rabbit All: all species expected

Trademarks and Patents

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

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Orders: 877-616-CELL (2355)•[email protected]• Support: 877-678-TECH (8324)•[email protected]• Web: cellsignal.com
仅供研究使用。不得用于诊断流程。
Revision 1
#7621

HTScan® SGK3 Kinase Assay Kit

HTScan® SGK3 Kinase Assay Kit: Image 1 Expand Image
图 1. 使用以下反应条件在放射测定法中测量 SGK3 激酶活性： 60 mM HEPES-NaOH、pH 7.5、3 mM MgCl2、3 mM MnCl2、3 µM 原钒酸钠、1.2 mM DTT、1 µM ATP、2.5 µg/50 µl PEG20,000，底物：GST-GSK3（14-27）：变量和重组 SGK3：变量。
HTScan® SGK3 Kinase Assay Kit: Image 2 Expand Image
图 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. 在 50 µl 反应体积中，每个反应孔在室温下用数量递增的 SGK3 和 1.5 µM 底物肽反应 30 分钟。（DELFIA® 是 PerkinElmer, Inc. 的注册商标）
HTScan® SGK3 Kinase Assay Kit: Image 3 Expand Image
图 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. 在 50 µl 反应体积中，每个反应孔在室温下用 50 ng SGK3、1.5 µM 底物肽、20 µM ATP 和数量递增的 staurosporine 反应 30 分钟。（DELFIA® 是 PerkinElmer, Inc. 的注册商标）
HTScan® SGK3 Kinase Assay Kit: Image 4 Expand Image
图 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. 在 50 µl 反应体积中，每个反应孔在室温下用 50 ng SGK3 和浓度递增的底物肽反应 30 分钟。（DELFIA® 是 PerkinElmer, Inc. 的注册商标）
HTScan® SGK3 Kinase Assay Kit: Image 5 Expand Image
图 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. 在 50 µl 反应体积中，每个反应孔用 50 ng SGK3 和 1.5 µM 底物肽。（DELFIA® 是 PerkinElmer, Inc. 的注册商标）
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Orders: 877-616-CELL (2355)•[email protected]• Support: 877-678-TECH (8324)•[email protected]• Web: cellsignal.com
仅供研究使用。不得用于诊断流程。