Autophagy assay cell type - Pancreata

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

CYTO-ID® Autophagy detection kit

Enzo Life Sciences

Upstream tips	Protocol tips	Downstream tips
Islets cells pretreated with rapamycin (500 nmol/L) can be used as positive control	After staining wash the cells with 1X assay buffer and transfer it to the glass slide for imaging. Add dye and incubate for 30 min at 37°C

Upstream tips
Islets cells pretreated with rapamycin (500 nmol/L) can be used as positive control
Protocol tips
After staining wash the cells with 1X assay buffer and transfer it to the glass slide for imaging. Add dye and incubate for 30 min at 37°C

Manufacturer protocol Publication protocol 1 Relevant paper

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