Autophagy Assay - Red Fluorescent

• Description:

  For the detection and monitoring the in vitro development of autophagy in living cells. The autophagy probe fluoresces red when inserted in the lipid membranes of autophagosomes and autolysosomes. Results can be read using a flow cytometer.

• Assay Principle:

  1. Prepare samples and controls., 2. Dilute 10X Cellular Assay Buffer 1:10 with diH2O., 3. Reconstitute FLICA with 50 µL DMSO., 4. Dilute FLICA 1:5 by adding 200 µL PBS., 5. Add diluted FLICA to each sample at 1:30-1:60 (e.g., spike at 1:30 by adding 10 µL to 290 µL sample)., 6. Incubate approximately 1 hour., 7. Remove media and wash cells 3 times: add 1X Cellular Wash Buffer and spin cells., 8. Resuspend cell pellet in 1X Cellular Wash Buffer., 9. If desired, label with additional stains, such as Hoechst 33342, DAPI, or an antibody., 10. If desired, fix cells., 11. Analyze with a fluorescence microscope or flow cytometer. FLICA 660 is excited at 660 nm and emits at 680-690 nm.

• Shipping Conditions:

  Blue Ice; Dry Ice

• Storage Conditions:

  4°C;-20°C