WB Solution Base Kit(Mouse)

To prepare the protein lysate for analysis, equal volumes of 2* loading buffer should be mixed in. The mixture should then be cooked at 100 ° C for 10 minutes and centrifuged at 12,000rpm for 5 minutes. Once done, take an appropriate volume to load the sample onto an electrophoresis system.
Next, the prepared electrophoresis solution should be used to carry out electrophoresis at 90V for 20 minutes followed by electrophoresis at 120V until the end. The next step involves film transfer for which an electrotransfer liquid should be prepared and used at a constant 100V for 90 minutes.
The subsequent step is closure, which entails incubating the film in a 5% skim milk powder/BSA solution with 1*TBST for 60 minutes on a shaking table at room temperature. Following this, incubate the primary antibodies overnight on a 2-8 degree shaking table.
To proceed, wash the film with 1*TBST for three 10-minute cycles. Thereafter, incubate the secondary antibody in a room temperature shaking table for 60 minutes. Afterwards, wash the film again with 1*TBST for three 10-minute cycles.
Finally, for color development and exposure, mix equal volumes of color development liquid A and liquid B and spread the mixed liquid over the whole film for 2 minutes. Only then can the exposure operation be carried out.