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.

数据：

LC3B-GFP fusion protein expressed and localized to autophagosomes

HeLa cells were transduced with Premo™ Autophagy sensor LC3B-GFP (green) and cultured overnight. The following day cells were nutrient deprived in Earles buffered salt solution containing 60uM Chloroquine. Images were acquired two hours after incubation with a Zeiss LSM 710 confocal microscope

Neonatal Human dermal fibroblast cells transduced with Premo™ Autophagy sensor LC3B-GFPNeonatal Human dermal fibroblasts (HDFn) cells were transduced with Premo™ Autophagy sensor LC3B-GFP and CellLight® Mito-RFP and cultured overnight in the presence of 100uM Chloroquine. The following day cells were counterstained with 1ug/mL Hoeschst 33342 (blue) and imaged on a Delta Vision wide field microscope.

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