eIF4B Antibody (YA3895)

Western blot analysis of extracts from Hela (lane 2(20μg), A549 (lane 3(20μg), HL-60 (lane 4(20μg), NIH/3T3 (lane 5(20μg) using eIF4B Antibody. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in TBST for 2 hour at room temperature. The primary antibody (1/1000)and Loading control antibody (Beta Actin, HY-P80438, 1/3000) was used in 5% BSA in TBST at 4°C overnight. Goat Anti-Mouse/Rabbit IgG-HRP Secondary Antibody (HY-P8004/HY-P8001, 1/10,000) was used for 1 hour at room temperature.

Flow cytometric analysis of 1X10⁶ HeLa cells labeling eIF4B Antibody (HY-P84198, red). Cells were fixed with 4% paraformaldehyde. Then stained with the primary antibody at 1/50 dilution for an hour at 4℃. AF 488-conjugated AffiniPure Goat Anti- Mouse IgG H&L (HY-P8005) was used as the secondary antibody at 1/1,000 dilution for 30 minutes at 4℃. Mouse IgG Isotype Control (HY-P80757, blue) was used as the isotype control, cells without incubation with primary antibody were used as the unlabeled control (black).

Immunocytochemistry analysis of HeLa cells labeling eIF4B with eIF4B Antibody (HY-P84198) at 1/500 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 quick block buffer for 10 minutes at room temperature. Cells were then incubated with eIF4B Antibody (HY-P84198) at 1/500 dilution in quick block buffer overnight at 4 ℃. AF488-conjugated Goat Anti-Mouse IgG H&L（HY-P8005, Green) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. The Nuclear counterstain was DAPI (Blue).

Immunocytochemistry analysis of NIH-3T3 cells labeling eIF4B with eIF4B Antibody (HY-P84198) at 1/500 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 quick block buffer for 10 minutes at room temperature. Cells were then incubated with eIF4B Antibody (HY-P84198) at 1/500 dilution in quick block buffer overnight at 4 ℃. AF488-conjugated Goat Anti-Mouse IgG H&L（HY-P8005, Green) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. The Nuclear counterstain was DAPI (Blue).