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The Lactate Dehydrogenase Sequestration Assay — A Simple and Reliable Method to Determine Bulk Autophagic Sequestration Activity in Mammalian Cells
JoVE Journal
Biology
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JoVE Journal Biology
The Lactate Dehydrogenase Sequestration Assay — A Simple and Reliable Method to Determine Bulk Autophagic Sequestration Activity in Mammalian Cells
DOI:

09:34 min

July 27, 2018

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Chapters

  • 00:04Title
  • 00:40Cell Seeding and Treatment
  • 02:46Cell Harvest and Preparation for Electrodisruption
  • 03:53Plasma Membrane Electrodisruption and Separation of Sedimentable- and Total Cell Fractions
  • 05:50LDH Extraction and Measurement of LDH Enzymatic Activity
  • 06:55Results: Monitoring Bulk Autophagic Sequestration Activity in Mammalian Cells
  • 08:43Conclusion

Summary

Automatic Translation

Here a simple and well-validated protocol for measuring bulk autophagic sequestration activity in mammalian cells is described. The method is based on quantifying the proportion of lactate dehydrogenase (LDH) in sedimentable cell fractions compared to total cellular LDH levels.

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