Autophagy assay cell type - RG2 [D74]

As autophagy is a multi-step process which includes not just the formation of autophagosomes, but most importantly, flux through the entire system, including the degradation upon fusion with lysosomes, which makes it quite challenging for detection. There are several methods for detection in mammalian cells, including immunoblotting analysis of LC3 and p62 and detection of autophagosome formation/maturation by fluorescence microscopy, Currently, there is no single “gold standard” for determining the autophagic activity that is applicable in every experimental context, hence it is recommended to go for the combined use of multiple methods to accurately assess the autophagic activity in any given biological setting.

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Found 1 matching solution for this experiment

LC3A (D50G8) XP® Rabbit mAb

Cell Signaling Technology

Upstream tips	Protocol tips	Downstream tips
	- Dilute Primary Ab at -1:1000. - For LC3 analyses the same PVDF-blotted membranes were cut horizontally at the 38 kDa marker so that the lower half could be blotted for LC3 while the upper half could be blotted simultaneously for actin to ensure equality of protein loading and transfer across samples.

Protocol tips
- Dilute Primary Ab at -1:1000. - For LC3 analyses the same PVDF-blotted membranes were cut horizontally at the 38 kDa marker so that the lower half could be blotted for LC3 while the upper half could be blotted simultaneously for actin to ensure equality of protein loading and transfer across samples.

Manufacturer protocol Publication protocol 1 Relevant paper

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