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The Lactate Dehydrogenase Sequestration Assay — A Simple and Reliable Method to Determine Bulk Autophagic Sequestration Activity in Mammalian Cells
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The Lactate Dehydrogenase Sequestration Assay — A Simple and Reliable Method to Determine Bulk Autophagic Sequestration Activity in Mammalian Cells

The Lactate Dehydrogenase Sequestration Assay — A Simple and Reliable Method to Determine Bulk Autophagic Sequestration Activity in Mammalian Cells

DOI:
10.3791/57971-v

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09:34 min

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July 27, 2018

•

, ,
1The Autophagy Team, Centre for Molecular Medicine Norway (NCMM), Nordic EMBL Partnership,University of Oslo

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

자동 번역

자동 번역

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.

Tags

Lactate Dehydrogenase Sequestration AssayAutophagyAutophagic SequestrationCell CultureTrypsinCell CountingCell SeedingBafilomycin A1Cell Detachment

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