Analysis of Autophagy by Flow Cytometry (CAT#: STEM-CBT-0059-WXH)

Flow cytometry has been used to detect the presence of autophagy mainly by the fluorescent antibody labeling of the autophagy marker, the microtubule associated protein LC3-II.
These techniques exploit the fact that LC3 protein, once associated with the inner autophagosomal membrane, is delivered into the lysosome and degraded there. Importantly, the GFP-tagged version is quenched prior to its degradation, owing to the sensitivity of this fluorescent protein to the acidic lysosomal environment. Accordingly, the reduction in GFP intensity in autophagy-induced samples relative to untreated samples serves as a readout for autophagic activity.

• Study a variety of pathophysiological processes.
• Study of autophagy-based therapies.

1.Induction of autophagy
2.Fixation and permeabilization
3.Labeling
4.Examine by flow cytometry

Autophagosome marker: light chain 3 (LC3)
Dyes: LysoTracker dyes, MitoTracker dyes