HTScan® TSK2 Kinase Assay Kit (#7630) Datasheet Without Images Cell Signaling Technology

For Research Use Only. Not for Use in Diagnostic Procedures.

UniProt ID:

#Q96PF2

Entrez-Gene Id:

23617

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.

Source / Purification

The GST-Kinase fusion protein was produced using a baculovirus expression system with a construct expressing full-length human TSK2 (Met1-Thr358) (GenBank Accession No. NM_053006) 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 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.

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).

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

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