Membranes were washed twice with TBS-T.
Membranes were washed three times with 0.1% TBS/Tween-20.
Subsequently membranes were washed with TBST and visualized using Odyssey imaging system (LI-COR).
Membranes were washed in the dark and then scanned using an Odyssey infrared imaging system (LI-COR Biosciences).
Following secondary antibody incubations, membranes were washed and finally incubated with ECL detection reagent (Amersham RPN2232) for 5 minutes.
The membranes were washed with TBS-T three times, and then subjected to immunoblotting using an anti-FLAG antibody.
The next day, membranes were washed with TBST and incubated with appropriate secondary antibodies for 1 hour at room temperature.
Then, the membranes were washed three times with TBS and incubated with appropriate secondary antibodies conjugated with horseradish peroxidase.
Thereafter, the membranes were washed with TTBS.
Membranes were washed for 30 min and incubated with horseradish peroxidase-conjugated secondary antibodies (Zhongshanjinqiao Corp).
The membranes were washed with 0.5 × SSC/0.1 % SDS at 55 °C.
