CYTO-ID® Autophagy detection kit

Protocol tips

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	The autophagy-related LC3 protein (ENZO) in cells was detected using Cyto-ID. Cells were grown on the coverslip and selected when the density was 50-70%, followed by intervention according to the experimental design; the negative control group was set up. The supernatant was discarded and the cells were washed twice with 1×Assay buffer for 30 min and were washed with 1×Assay buffer, and the excess buffer was removed, followed by sealing using anti-quenching agent containing 4’6-diamidino-2-phenylindole (DAPI) dye. Then, cells were incubated in the dark at 37ºC . The coverslip was observed under fluorescence microscope (FITC and DAPI; 60×).

Protocol tips

The autophagy-related LC3 protein (ENZO) in cells was detected using Cyto-ID. Cells were grown on the coverslip and selected when the density was 50-70%, followed by intervention according to the experimental design; the negative control group was set up. The supernatant was discarded and the cells were washed twice with 1×Assay buffer for 30 min and were washed with 1×Assay buffer, and the excess buffer was removed, followed by sealing using anti-quenching agent containing 4’6-diamidino-2-phenylindole (DAPI) dye. Then, cells were incubated in the dark at 37ºC . The coverslip was observed under fluorescence microscope (FITC and DAPI; 60×).

Publication protocol

The autophagy-related LC3 protein (ENZO) in cells
was detected using Cyto-ID. Cells were grown on the
coverslip and selected when the density was 50-70%,
followed by intervention according to the experimental design; the negative control group was set up. The
supernatant was discarded and the cells were washed
twice with 1×Assay buffer for 30 min and were washed
with 1×Assay buffer, and the excess buffer was removed,
followed by sealing using anti-quenching agent containing 4’6-diamidino-2-phenylindole (DAPI) dye. Then, cells were incubated in the dark at 37ºC . The coverslip was observed under fluorescence microscope (FITC and DAPI; 60×).

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