Enzo Life Sciences Cyto-ID Autophagy Detection Kit

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The Cyto-ID Autophagy Detection Kit from Enzo Life Sciences measures autophagic vacuoles and monitors autophagic flux in live cells using a novel dye that selectively labels autophagic vacuoles. The dye has been optimised 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 assay offers a rapid and quantitative approach to monitoring autophagy in live cells without the need for cell transfection.

Key advantages include: no transfection required; dye includes titratable moieties specific for selectively staining autophagic vesicles; protocol validated with known inhibitors and activators of autophagic activity; rapidly quantifies autophagy in native heterogeneous cell populations; eliminates need for time- and effort-consuming transfection efficiency validation required with LC3-GFP transfection; selective and comprehensive staining allows measurement and differentiation between autophagic flux and autophagolysosome accumulation; negligible staining of lysosomes reduces background seen with other dyes; facilitates high-throughput screening of activators and inhibitors of autophagy.

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 labelling of vacuoles associated with the autophagy pathway.

The kit is a member of the CELLestial product line, reagents and assay kits comprising fluorescent molecular probes that have been extensively benchmarked for live cell analysis applications.