LC3B is one of the mammalian Atg8 homologs and widely used as an autophagosome marker. Immediately after synthesis, LC3 is processed by Atg4 and becomes LC3-I. Upon induction of autophagy, the C-terminal glycine of LC3-I is conjugated to phosphatidylethanolamine, resulting in formation of membrane-bound LC3-II. Most LC3-II is thought be present on autophagosome membrane. The autophagosome subsequently fuses with a lysosome, where inside materials, including LC3-II, are degraded. The expression level of LC3-II generally correlates with the number of autophagosome
Western Blot: 1/500-1/1,000.
Predicted molecular weight: LC3-I is 18 kDa and LC3-II is 16 kDa
MEF +/- Starvation 2 hr
SDS-PAGE (MEF lysate 15 µ/lane, 15% gel)
Transfer to PVDF membrane (semi-dry blotter, 100 mA/gel 1 hr)
Immunoblot for LC3B
Expose ECL 5min
Optimum dilution rate should be determined by end users
To be used for research only. DO NOT use for human gene therapy or clinical diagnosis.