Lentivirus express a fusion of “RFP-LC3″. The codon sequence of human LC3 gene (gene_synonym: ATG8F; LC3B; MAP1A/1BLC3; MAP1LC3B-a; NCBI ID: NM_022818 ), was fusioned in frame with Red Fluorescent Protein (RFP) at its N-terminal, expressed under a constitutive suCMV promoter, containing a puromycin antibiotic marker under the RSV promoter.
By creating fluorescent-LC3 fusion constructs, you can visualize autophagy in real time in live cells under fluorescent-microscope. Also, the turnover of Fluorescent-LC3 can be quantify using flow cytometry to measure autophagic activity in living mammalian cells.
During autophagosome formation, the fusion protein is processed and recruited to the autophagosome membrane, where it can be imaged as cytoplasmic puncta by high resolution fluorescence microscopy. The changes in percentage of Fluorescent positive cells can be determined and serves as the indicative of autophagy processing stages. see Product Manual for all fusion products.
Amount: 200ul/vial (1×107 IFU/ml), provided in DMEM medium, containing 60 ug/ml of Polybrene (10x).