Imagequant las 54000 imaging system software

Protein samples were analyzed by 10% and 12.5% SDS-PAGE (65 ). Proteins were transferred from the gels to polyvinylidene difluoride (PVDF) membranes using the Trans-Blot Turbo system (Bio-Rad). Western blot analysis was performed with a monoclonal antibody raised against the Flag epitope (Sigma) diluted 1:10,000 in a solution containing phosphate-buffered saline (PBS), 0.2% Triton, and 3% skim milk and incubated for 16 h at 4°C. The membranes were washed for 20 min each with PBS and 0.2% Triton solution. The washing step was repeated three times. Thereafter, the membranes were incubated with horseradish peroxidase-conjugated goat anti-mouse IgG (Invitrogen) diluted 1:5,000 in a solution containing PBS and 0.2% Triton for 1 h at room temperature. Again, three washing steps of 45 min with PBS and 0.2% Triton solution were performed, and detection was performed by enhanced chemiluminescence using ImageQuant LAS 54000 imaging system software (GE Healthcare Lifesciences).

Protein samples were analyzed by SDS-PAGE with 10% or 12.5% gels [68 (link)]. Proteins were transferred from the gels to PVDF membranes using the Trans-Blot Turbo system (Bio-rad). Western blot analysis was performed with a monoclonal antibody raised against the Flag-epitope (Sigma) diluted 1:10,000 in a solution of PBS, 0.2% Triton, 3% skimmed milk and incubated for 16 hours at 4°C. Membranes were washed for 20 minutes each with PBS, 0.2% Triton solution. The washing step was repeated three times. Thereafter, the membranes were incubated with horseradish peroxidase-conjugated goat anti-mouse IgG (Promega) diluted 1:2500 in a solution of PBS, 0.2% Triton for 1 hour at room temperature. Again, three washing steps of 45 minutes with PBS, 0.2% Triton solution were performed, and detection was performed by enhanced chemiluminescence using ImageQuant LAS54000 imaging system software (GE Healthcare Lifesciences).