Western blot analysis of LC3B on different lysates with Rabbit anti-LC3B antibody (ET1701-65) at 1/2;000 dilution and competitor's antibody at 1/1;000 dilution. Lane 1: HeLa cell lysate Lane 2: HeLa treated with 50μM Chloroquine for 18 hours cell lysate Lane 3: C2C12 cell lysate Lane 4: C2C12 treated with 50μM Chloroquine for 18 hours cell lysate Lane 5: C6 cell lysate Lane 6: C6 treated with 50μM Chloroquine for 18 hours cell lysate Lysates/proteins at 20 µg/Lane. Predicted band size: 14/16 kDa Observed band size: 14/16 kDa Exposure time: 3 minutes; 4-20% SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1701-65) at 1/2;000 dilution and competitor's antibody at 1/1;000 dilution were used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50;000 dilution was used for 1 hour at room temperature.
Western blot analysis of ATG5 on different lysates with Rabbit anti-ATG5 antibody (ET1611-38) at 1/5;000 dilution and competitor's antibody at 1/1;000 dilution. Lane 1: NIH/3T3 cell lysate (15 µg/Lane) Lane 2: C2C12 cell lysate (15 µg/Lane) Lane 3: Neuro-2a cell lysate (15 µg/Lane) Lane 4: PC-12 cell lysate (15 µg/Lane) Lane 5: Mouse brain tissue lysate (15 µg/Lane) Lane 6: Rat brain tissue lysate (15 µg/Lane) Predicted band size: 32 kDa Observed band size: 55 kDa Exposure time: 35 seconds; 4-20% SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1611-38) at 1/5;000 dilution and competitor's antibody at 1/1;000 dilution were used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50;000 dilution was used for 1 hour at room temperature.