CYTO-ID^® Autophagy detection kit 2.0

• Brighter, more photostable dye specifically stains autophagic vesicles
• Rapid, no transfection required
• Negligible staining of lysosomes reduces background seen with other dyes
• Facilitates high-throughput screening of activators and inhibitors of autophagy
• Widely cited

Enzo Life Sciences CYTO-ID^® Autophagy Detection Kit 2.0 measures autophagic vacuoles and monitors autophagic flux in lysosomally inhibited live cells using a novel dye that selectively labels accumulated autophagic vacuoles. The 488nm-excitable green dye has been optimized through the identification of titratable functional moieties that allow for minimal staining of lysosomes while exhibiting bright fluorescence upon incorporation into pre-autophagosomes, autophagosomes, and autolysosomes (autophagolysosomes). The kit also includes the Hoechst 33342 dye for the nuclear staining, an Autophagy Inducer (Rapamycin) and a Lysosomal Inhibitor (Chloroquine).

Mechanism of Action
The probe is a cationic amphiphilic tracer (CAT) dye that rapidly partitions into cells in a similar manner as drugs that induce phospholipidosis. Careful selection of titratable functional moieties on the dye prevents its accumulation within lysosomes, but enables labeling of vacuoles associated with the autophagy pathway.

Autophagy is a stress-induced protective mechanism. Less active under basal conditions, the mechanism is utilized by eukaryotic cells through lysosome-mediated bulk degradation of cellular contents when subjected to certain hostile conditions such as nutrient depletion and chemical or environmental stress. The role of increased autophagic activity in the pathology of cancer, neurodegeneration, cardiovascular disease and diabetes has become widely recognized and commonly studied. Induction of autophagic flux can be visualized by enhanced accumulation of autophagic vesicles if lysosomal function is inhibited, preventing removal of these vesicles.