ETFA Antibody (YA3789)

Western blot analysis of extracts from MCF-7(lane 1(40μg) ),Hela(lane 2(40μg) ),HEK293(lane 3(40μg) ),HepG2(lane 4(40μg) )and A431(lane 5(40μg) ) using ETFA antibody. Proteins were transferred to a NC membrane and blocked with 5% Skim milk in TBST for 2 hour at room temperature. The primary antibody ( 1/1000) and Loading control antibody (GAPDH，1/1000) was used in 5% Skim milk in TBST at 4°C overnight. Goat Anti-Mouse IgG-HRP Secondary Antibody (1/10,000) was used for 1 hour at room temperature.

Flow cytometric analysis of 1X10^6 Hela cells labeling ETFA Antibody(red). Cells were fixed with 4% paraformaldehyde and permeabilised with 0.2% Triton X-100. Then stained with the primary antibody at 1/400 dilution overnight at 4℃. AF 488 Goat Anti-mouse IgG H&L was used as the secondary antibody at 1/1,000 dilution for 45 minutes at room temperature. Mouse IgG Isotype Control (blue) was used as the isotype control, cells without incubation with primary antibody were used as the unlabeled control (black).

Immunofluorescence analysis of A549 cells labeling ETFA antibody at 1/50 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature.Cells were then incubated with ETFA antibody at 1/50 dilution in 1% BSA in PBST overnight at 4 ℃. AF 488 Goat Anti-mouse IgG H&L (green) was used as the secondary antibody at 1/500 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. The Nuclear counterstain was DAPI (Blue)

Immunofluorescence analysis of Sk-Br-3 cells labeling ETFA antibody at 1/50 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature.Cells were then incubated with ETFA antibody at 1/50 dilution in 1% BSA in PBST overnight at 4 ℃. AF 488 Goat Anti-mouse IgG H&L (green) was used as the secondary antibody at 1/500 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. The Nuclear counterstain was DAPI (Blue)