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Characterization at the Molecular Level using Robust Biochemical Approaches of a New Kinase Protein
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Biochemistry
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JoVE Journal Biochemistry
Characterization at the Molecular Level using Robust Biochemical Approaches of a New Kinase Protein

Characterization at the Molecular Level using Robust Biochemical Approaches of a New Kinase Protein

DOI:
10.3791/59820-v

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11:23 min

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June 30, 2019

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, , ,
1Centre de Biophysique Moléculaire, CNRS, UPR 4301,University of Orléans and INSERM

Chapters

  • 00:04Title
  • 01:24Transient Transfections
  • 03:21Lysis
  • 05:17Immunoprecipitation
  • 06:59Coimmunoprecipitation Analyses
  • 08:33Results: Synthesized LIMK2-1 Protein, Its Interaction with Kinase ROCK, its Kinase Activity
  • 10:10Conclusion

Summary

Automatic Translation

Automatic Translation

We characterized a new kinase protein using robust biochemical approaches: Western Blot analysis with a dedicated specific antibody on different cell lines and tissues, interactions by coimmunoprecipitation experiments, kinase activity detected by Western Blot using a phospho-specific antibody and by γ[32P] ATP labeling.

Tags

Keywords: ImmunoprecipitationKinaseProtein ActivitySubstrateP32-ATP LabelingInhibitorLysis ConditionsNegative ControlCell CultureTransfectionCalcium Phosphate

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