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Apoptosis pp 161-172 | Cite as

Measurement of Changes in Cdk2 and Cyclin O-Associated Kinase Activity in Apoptosis

Protocol
First Online:
Part of the Methods in Molecular Biology book series (MIMB, volume 559)

Summary

Many cell cycle regulatory proteins have been shown to be able to regulate cell death. Activation of Cdk2 has been shown to be necessary for apoptosis of quiescent cells such as thymocytes, neurons, and endothelial cells. This activation is stimulus-specific because it occurs in glucocorticoid and DNA damage but not in CD95-induced apoptosis in thymocytes. Apoptotic Cdk2 activation in lymphoid cells is controlled by a recently identified protein, cyclin O, and its activity is modulated by p53 and members of the Bcl-2 protein family. In this chapter, we describe methods for measuring changes in Cdk2 activity during apoptosis. In addition, we also show the details of the generation of an antibody able to immunoprecipitate the cyclin O complexes from apoptotic cells in native conditions and its use to measure the kinase activity associated with this proapoptotic cyclin.

Key words

Cdk2 Apoptosis Thymocytes Cell cycle Kinase Intrinsic stimuli 
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Copyright information

© Humana Press, a part of Springer Science+Business Media, LLC 2009

Authors and Affiliations

  1. 1.Institut Municipal d’Investigació Mèdica (IMIM)BarcelonaSpain

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