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7630
HTScan® TSK2 Kinase Assay Kit
Cellular Assay Kits

HTScan® TSK2 Kinase Assay Kit #7630

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Figure 3. Dose dependence curve of TSK2 kinase activity: DELFIA® data generated using Phospho-PLK (Ser137) Antibody #5070 to detect phosphorylation of substrate peptide (#1300) by TSK2 kinase. In a 50 µl reaction, increasing amounts of TSK2 and 1.5 µM substrate peptide were used per reaction at room temperature for 30 minutes. (DELFIA® is a registered trademark of PerkinElmer, Inc.)

Figure 5. Staurosporine inhibition of TSK2 kinase activity: DELFIA® data generated using Phospho-PLK (Ser137) Antibody #5070 to detect phosphorylation of TSK2 substrate peptide (#1300) by TSK2 kinase. In a 50 µl reaction, 50 ng TSK2, 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.)

Figure 4. Peptide concentration dependence of TSK2 kinase activity: DELFIA® data generated using Phospho-PLK (Ser137) Antibody #5070 to detect phosphorylation of substrate peptide (#1300) by TSK2 kinase. In a 50 µl reaction, 50 ng of TSK2 and increasing concentrations of substrate peptide were used per reaction at room temperature for 30 minutes. (DELFIA® is a registered trademark of PerkinElmer, Inc.)

Figure 2. Time course of TSK2 kinase activity: DELFIA® data generated using Phospho-PLK (Ser137) Antibody #5070 to detect phosphorylation of TSK2 substrate peptide (#1300) by TSK2 kinase. In a 50 µl reaction, 50 ng TSK2 and 1.5 µM substrate peptide were used per reaction. (DELFIA® is a registered trademark of PerkinElmer, Inc.)

Figure 1. TSK2 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: Histone H2A, variable, and recombinant TSK2: variable.

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Product Description

The kit provides a means of performing kinase activity assays with recombinant human TSK2 kinase. It includes active TSK2 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-peptide: 1,646 Daltons. GST-TSK2 Kinase domain: 74 kDa.

Peptide Core Sequence:

RRS*LL

Background

Testis-specific kinases (TSKs) are serine/threonine kinases that may play a role in sperm function. Three TSKs, TSK1, TSK2 and TSK3 have been identified. Open reading frames (ORF) within the TSK gene code for the 3 isoforms: 367 amino acids for TSK1, 358 amino acids for TSK2 and 275 amino acids for TSK3. The kinase domains are located near the NH2-terminal regions of the TSK proteins and contain conserved amino acid residues characteristic of a serine/threonine kinase. While TSK1 and TSK2 both have a 100 amino acid domain located C-terminally to the kinase domain, TSK3 is composed essentially of a serine/threonine kinase domain. TSK1 and TSK2 messenger RNA is expressed exclusively in the testis of mature males. An in vitro kinase assay of immunoprecipitates containing either TSK1 or TSK2 revealed serine phosphorylation of a coprecipitating protein of approximately 65 kDa. This substrate protein is also expressed exclusively in the testis as demonstrated by northern and dot blot analyses (1,2). While TSK1 and TSK2 form detergent resistant complexes during immunoprecipitation, TSK3 does not associate with either protein. Expression of TSK3 is induced at puberty, is restricted to the interstitial Leydig cells of postpubertal males, persists throughout adulthood and is implicated in regulating androgenesis and/or Leydig cell proliferation (4).

  1. Bielke, W. et al. (1994) Gene 139, 235-239.
  2. Hao, Z. et al. (2004) Mol Hum Reprod 10, 433-444.
  3. Kueng, P. et al. (1997) J Cell Biol 139, 1851-1859.
  4. Zuercher, G. et al. (2000) Mech Dev 93, 175-177.

Pathways & Proteins

Explore pathways + proteins related to this product.

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.