Autophagy Assay Kit

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	- Add autophagosome reagent incubate for 1h at 37 °C, 5% CO2

Protocol tips
- Add autophagosome reagent incubate for 1h at 37 °C, 5% CO2

Publication protocol

The effect of NPs, formulated with or without CPP, on inducing autophagy was assessed using Autophagy Assay Kit (Sigma, Castle Hill, NSW, Australia), as per manufacturer’s instructions. Briefly, cells (A549 and Beas-2B) were seeded using complete media in a black clear bottom 96-well plate (Corning, Sigma) at 2 × 104 cells/well and allowed to attach and grow for 24 h. Cells were exposed to the NPs, formulated with or without CPP for 24 h. Treatments were removed and the supplied autophagosome reagent was added to each well and incubated for 1h at 37 °C, 5% CO2. Cells were carefully washed 4 times with washing buffer provided in the kit and fluorescence signal measured at 360/520 nm using a spectrophotometer. NPs–DNA formulated or not with CPP were tested in a concentration range from 0.4 to 2.4 mg/mL of PLGA. CPP alone was also tested at different concentrations. Results are presented in relation to the media control signal and were analysed using one-way ANOVA, means were compared using Tukey’s test.

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 - Beas-2B using Autophagy Assay Kit from Sigma-Aldrich.

Manufacturer protocol

Download the product protocol from Sigma-Aldrich for Autophagy Assay Kit below.

Download manufacturer protocol

Videos

Check out videos that might be relevant for performing Autophagy assay cell type - Beas-2B using Autophagy Assay Kit from Sigma-Aldrich. Please note that these videos are representative and steps or experiment specific processes must be kept in mind to expect desired results.