Muse™Autophagy LC3-antibody based Kit

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	- Stain on ice for 30 minutes in the dark after adding Anti-LC3.	- The kit is designed to measure endogenous levels of LC3.

Protocol tips
- Stain on ice for 30 minutes in the dark after adding Anti-LC3.
Downstream tips
- The kit is designed to measure endogenous levels of LC3.

Publication protocol

Osteosarcoma 143B cells were seeded onto 10-cm culture dishes at a density of 4×104 cells/dish. After 24 h, the cells were incubated with 2-ME and/or L-lactate according to the experimental design. The cells were then pelleted and incubated with antibody detection reagents and stained with Anti-LC3 Alexa Fluor® 555, clone 4E12 according to manufacturer’s protocol. Afterward, the cells were analyzed (5,000 events/sample), and the signals were detected using the Muse Cell Analyzer (Merck Millipore, Poland). The results were then analyzed with the Muse 1.4 analysis software. Each experiment was performed at least three times.

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Manufacturer protocol

Download the product protocol from Merck Millipore for Muse™Autophagy LC3-antibody based Kit below.

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