Western blot analysis of Clathrin heavy chain on different lysates with Rabbit anti-Clathrin heavy chain antibody (ET1704-50) at 1/1;000 dilution. Lane 1: HeLa cell lysate Lane 2: A431 cell lysate Lane 3: NIH/3T3 cell lysate Lane 4: PC-12 cell lysate Lane 5: HUVEC cell lysate Lane 6: SH-SY5Y cell lysate Lane 7: COS-1 cell lysate Lysates/proteins at 20 µg/Lane. Predicted band size: 180kDa Observed band size: 180 kDa Exposure time: 10 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 (ET1704-50) at 1/1;000 dilution was 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 Caveolin-1 on different lysates with Rabbit anti-Caveolin-1 antibody (ET1603-1) at different dilutions. Lane 1/2: A549 cell lysate Lane 3/4: A431 cell lysate Lysates/proteins at 10 µg/Lane. Predicted band size: 20 kDa Observed band size: 20 kDa Exposure time: 2 minutes; 12% 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 (ET1603-1) at different dilutions were used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:300;000 dilution was used for 1 hour at room temperature.
All lanes: Western blot analysis of RAB7 with anti-RAB7 antibody (ET1611-96) at 1:500 dilution. Lane 1: Wild-type Hela whole cell lysate (10 µg). Lane 2/3: RAB7 knockdown Hela whole cell lysate (10 µg). ET1611-96 was shown to specifically react with RAB7 in wild-type Hela cells. Weakened bands were observed when RAB7 knockdown samples were tested. Wild-type and RAB7 knockdown samples were subjected to SDS-PAGE. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM in TBST for 1 hour at room temperature. The primary antibody (ET1611-96; 1:500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG-HRP Secondary Antibody (HA1001) at 1:300;000 dilution was used for 1 hour at room temperature.
Western blot analysis of RAB7 on different lysates with Rabbit anti-RAB7 antibody (ET1611-96) at 1/5;000 dilution. Lane 1: HeLa cell lysate Lane 2: MCF7 cell lysate Lane 3: Neuro-2a cell lysate Lane 4: C2C12 cell lysate Lane 5: PC-12 cell lysate Lane 6: C6 cell lysate Lysates/proteins at 15 µg/Lane. Predicted band size: 24 kDa Observed band size: 24 kDa Exposure time: 17 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 (ET1611-96) at 1/5;000 dilution was 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.
