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7555
HTScan® NEK2 Kinase Assay Kit
Cellular Assay Kits
Assay Kit

HTScan® NEK2 Kinase Assay Kit #7555

Citations (0)
HTScan® NEK2 Kinase Assay Kit: Image 1
Western blot analysis of extracts from serum-starved A431 cells, phosphorylated in vitro with PKA kinase or treated in culture with forskolin/IBMX, using Phospho-PKA Substrate (RRXS*/T*) (100G7E) Rabbit mAb. Lysis buffer: 1.0% Triton X-100 (lanes 1 and 2), 2.0% SDS (lanes 3 and 4).
Inquiry Info.# 7555

Product Description

The kit provides a means of performing kinase activity assays with recombinant human NEK2 kinase. It includes full-length human active NEK2 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-CREB (Ser133) peptide: 2,326 Daltons. GST-NEK2 Kinase: 83 kDa.

Background

The NEK family of protein kinases is composed of 11 members in humans that share an amino-terminal catalytic domain related to NIMA, a serine/threonine kinase identified in Aspergillus nidulans. While NIMA is critical for cell cycle progression in fungus, the function of NEK kinases in mammalian cells is largely unknown. NEK1 was first identified by screening mouse cDNA expression libraries and was demonstrated to have dual specificity kinase activity on both tyrosine and serine/threonine sites (1). NEK2 most closely resembles fungal NIMA in its primary structure and is believed to promote the splitting of duplicated centrosomes at the onset of mitosis (2,3). NEK3 is predominantly a cytoplasmic enzyme and its activity shows marginal variation throughout the cell cycle (4). NEK4 is ubiquitously expressed and its expression and subcellular location are not associated with cell cycle (5). NEK6/7 have been suggested to phosphorylate and activate p70 S6 kinase in vitro (6). Expression of an inactive NEK6 mutant arrests cells in M phase and interferes with chromosome segregation (7). NEK8 activity is not cell cycle regulated and may play a role in cell cycle independent microtubule dynamics (8). NEK9 is activated during mitosis and may participate in the activation of NEK6/7 during mitosis (9,10).
  1. Letwin, K. et al. (1992) EMBO J. 11, 3521-3531.
  2. Fry, A.M. et al. (1995) J. Biol. Chem. 270, 12899-12905.
  3. Fry, A.M. (2002) Oncogene 21, 6184-6194.
  4. Tanaka, K. and Nigg, E.A. (1999) J. Biol. Chem. 274, 13491-13497.
  5. Hayashi, K. et al. (1999) Biochem. Biophys. Res. Commun. 264, 449-456.
  6. Belham, C. et al. (2001) Curr. Biol. 11, 1155-1167.
  7. Yin, M.J. et al. (2003) J. Biol. Chem. 278, 52454-52460.
  8. Holland, P.M. et al. (2002) J. Biol. Chem. 277, 16229-16240.
  9. Belham, C. et al. (2003) J. Biol. Chem. 278, 34897-34909.
  10. Roig, J. et al. (2002) Genes Dev. 16, 1640-1658.

Pathways & Proteins

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