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Experimental Approaches to Study Mitochondrial Localization and Function of a Nuclear Cell Cycle Kinase, Cdk1
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Experimental Approaches to Study Mitochondrial Localization and Function of a Nuclear Cell Cycle Kinase, Cdk1

Experimental Approaches to Study Mitochondrial Localization and Function of a Nuclear Cell Cycle Kinase, Cdk1

DOI:
10.3791/53417-v

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13:15 min

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February 25, 2016

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, , ,
1Radiation Oncology,University of California, Davis

Chapters

  • 00:05Title
  • 01:01Sodium Carbonate Extraction of Intact Mitochondria
  • 03:07Separation of Inner and Outer Membranes of Mitochondria
  • 04:22Construction of Mitochondria-targeted GFP/RFP-tagged CyclinB1/Cdk1 Vectors and Mitochondria Localization
  • 07:23Determination of Cell Cycle Phase Lengths with EdU Incorporation Assay
  • 10:58Results: Mitochondrial Analysis of CyclinB1 and Cdk1
  • 12:25Conclusion

Summary

Automatic Translation

Automatic Translation

Here, we outline how to study mitochondrial localization of a (cell cycle) kinase, and how to determine its sub-mitochondrial location as well as potential mitochondrial substrates/targets. Forced expression of proteins into the mitochondria provides a useful tool for studying the functional consequences of mitochondrial localization of a protein of interest.

Tags

Keywords: Mitochondrial LocalizationNuclear Cell Cycle KinaseCdk1Mitochondrial ResearchProtein TaggingSubcellular FractionationSodium Carbonate ExtractionTrichloroacetic Acid PrecipitationImmunoblotting

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