Cell Harvest and Preparation for Electrodisruption
3:53
Plasma Membrane Electrodisruption and Separation of Sedimentable- and Total Cell Fractions
5:50
LDH Extraction and Measurement of LDH Enzymatic Activity
6:55
Results: Monitoring Bulk Autophagic Sequestration Activity in Mammalian Cells
8:43
Conclusion
Transcript
This method can help answer key questions in the autophagy field, such as how autophagosome formation is regulated and how various cell treatments and conditions affect bulk autophogy. The main advantage for this technique is that it measures endo
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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.