FlowCellect Autophagy LC3 antibody based kit

Protocol tips

Upstream tips	Protocol tips	Downstream tips
This Kit provides a quantitative solution for studying autophagy & measuring potency of autophagy inducers using flow cytometry.		If there is loss or lack of signal increase the cell number

Upstream tips
This Kit provides a quantitative solution for studying autophagy & measuring potency of autophagy inducers using flow cytometry.
Downstream tips
If there is loss or lack of signal increase the cell number

Publication protocol

Detection of autophagic punctas was performed as previously described [18]. Briefly, treated Hela cells were seeded on sterile coverslips and incubated under the conditions mentioned above. The growth medium was removed and cells were washed with cold PBS for two times when the cells’ confluence reached 80%. Then, fresh growth medium was supplemented subsequently and cells were incubated for an additional 12 h. Detection of GFP–LC3 was performed using FlowCellect™ GFP–LC3 Reporter Autophagy Assay kit (Millipore, U.S.A.). Briefly, 10 μl autophagy reagent A was added and cells were incubated in a humidified incubator at 37°C with 5% CO2 for 2 h. Post the removal of medium, cells were washed with 5 ml 1× HBSS, were added with 100 ml 1× autophagy reagent B, and then were cultured with cells for 5 min, followed by washing with assay buffer for the autophagy reagent removal. Last, the coverslips were covered by sterile slides and the slides were observed under an Olympus fluorescence microscope (BX51, Olympus Corporation, Japan).

Full paper   Login or join for free to view the full paper.

Reviews

Discussion

Papers

Check out relevant papers found by Labettor's AI that are relevant for performing Autophagy assay cell type - HeLa using FlowCellect Autophagy LC3 antibody based kit from Millipore.

Manufacturer protocol

Download the product protocol from Millipore for FlowCellect Autophagy LC3 antibody based kit below.

Download manufacturer protocol

Videos

Check out videos that might be relevant for performing Autophagy assay cell type - HeLa using FlowCellect Autophagy LC3 antibody based kit from Millipore. Please note that these videos are representative and steps or experiment specific processes must be kept in mind to expect desired results.