Autophagy assay cell type - MRC5

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 3 matching solutions for this experiment

Upstream tips
This dual staining autophagy/cytoxicity cell-based assay facilitates the detection of both cells showing indicators of autophagy and cells showing indicators of cytotoxicity.
Dansylcadaverine

Sigma-Aldrich

Upstream tips
MDC is a fluorescent compound that stains autophagic vacuoles.
Protocol tips
Treat the cells with 0.05mM MDC for 1hour at 37°C
Beclin-1 Antibody

Cell Signaling Technology

Protocol tips
Primary Ab dilution-1:1000 dilution
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