African Journal of
Biotechnology

  • Abbreviation: Afr. J. Biotechnol.
  • Language: English
  • ISSN: 1684-5315
  • DOI: 10.5897/AJB
  • Start Year: 2002
  • Published Articles: 12200

Full Length Research Paper

Monitoring cyclin-dependent kinase 2 (CDK2) activity with Kinase-Glo, a luminescence-based ATP quantitative reagent

Qi Liu, Jinlan Gao, Xing Liu, Shusen Wang and Yang Luo*
The Research Center for Medical Genomics and MOH Key Laboratory of Cell Biology, China Medical University, Shenyang 110001, China.
Email: [email protected]

  •  Accepted: 03 January 2011
  •  Published: 14 February 2011

Abstract

 

Protocols assessing kinase activity generally include radioactive methods, fluorescent polarization and the use of anti-phosphorylation antibodies. The present study described the application of a luminescence-based ATP quantitative reagent, Kinase-Glo, in evaluating the activity of cyclin-dependent kinase 2 (CDK2). Different amounts of immunoprecipitated CDK2 retrieved from HeLa extracts and different concentrations of the CDK2 inhibitor, roscovitine, were applied to test the sensitivity of Kinase-Glo reagent. The results demonstrated that Kinase-Glo reagent was sensitive to detect the alterations of CDK2 activity and suggested it to be applied in assessing kinase activity and for inhibitor optimization in vitro.

 

Key words: Cyclin-dependent kinase 2 (CDK2), kinase assay, Kinase-Glo, relative light unit (RLU), luminescence.

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