Premo™ Autophagy Sensor combines the selectivity of a LC3B-green fluorescent protein (GFP) chimera with the transduction efficiency of BacMam technology, enabling unambiguous visualization of this protein. BacMam reagents (insect Baculovirus with a Mammalian promoter) are non-replicating in mammalian cells and thus safe to handle. They are also non-cytotoxic and ready-to-use. Unlike expression vectors, BacMam reagents enable titratable and reproducible expression and offer high co-transduction efficiency, therefore multiple BacMam reagents can readily be used in the same cell. Recent improvements made to the BacMam system enable efficient transduction in a wider variety of cells including neurons and neural stem cells (NSCs) with an easy, one-step protocol. Now, to visualize autophagy, simply add the BacMam LC3B-GFP to the cells and incubate overnight for protein expression. Each Premo™ Autophagy Sensor Kit includes the BacMam LC3B-FP, a control BacMam LC3B (G120A)-FP (mutation on the control BacMam LC3B prevents cleavage and subsequent lipidation during normal autophagy, thus protein localization should remain cytosolic and diffuse) and chloroquine diphosphate to artificially induce autophagosome accumulation.
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