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Study of the Functions and Activities of Neuronal K-Cl Co-Transporter KCC2 Using Western Blotting
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JoVE Journal Biyokimya
Study of the Functions and Activities of Neuronal K-Cl Co-Transporter KCC2 Using Western Blotting

Study of the Functions and Activities of Neuronal K-Cl Co-Transporter KCC2 Using Western Blotting

DOI:
10.3791/64179-v

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10:08 min

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December 09, 2022

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, , ,
1Institute of Biomedical and Clinical Sciences, Medical School, Faculty of Health and Life Sciences,University of Exeter, Hatherly Laboratories

Bölümler

  • 00:05Introduction
  • 00:50Cell Culture and Transfection
  • 02:28Preparation of Cell Lysates and Loading Samples
  • 03:11Preparation of Immunoprecipitants from Cell Lysates
  • 04:40Performing the Western Blot
  • 06:23Antibody Staining, Image Development, Image Acquisition, and Data Quantification
  • 07:16Results: Western Blotting Technique to Study the Functions and Activities of Neuronal K-Cl Co-Transporter KCC2
  • 09:25Conclusion

Özet

Otomatik Çeviri

Otomatik Çeviri

The present protocol highlights the application of western blotting technique to study the functions and activities of neuronal K-Cl co-transporter KCC2. The protocol describes the investigation of KCC2 phosphorylation at kinase regulatory sites Thr906/1007 via western blotting. Also, additional methods to confirm KCC2 activity are briefly highlighted in this text.

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