WB Solution Base Kit(Goat)

Here are the steps for protein lysate preparation and analysis:
1. Mix the protein lysate with 2* loading buffer in equal volume and heat it at 100 ° C for 10min. Then, centrifuge it at 12000rpm for 5min and take an appropriate volume to load the sample.
2. Electrophoresis can be performed using the prepared electrophoresis solution. Electrophoresis at 90V for 20min, and continue electrophoresis at 120V until the end.
3. Film transfer is the next step. Use the prepared electrotransfer liquid and apply a constant piezoelectric of 100V for 90min.
4. The film is then placed in a 5% skim milk powder/BSA solution with 1*TBST. Incubate for 60min on a shaking table at room temperature to prepare it for antibody incubation.
5. Incubate primary antibodies overnight on a 2-8 degree shaking table.
6. Wash the film with 1*TBST for three times, with each wash taking 10min.
7. Incubate the secondary antibody in a room temperature shaking table for 60min.
8. Repeat the washing process three times, with each wash taking 10min.
9. The final step is color development and exposure. Mix color development liquid A and liquid B in equal volume and apply the mixed liquid over the entire film for 2min. Finally, carry out the exposure operation.
By following these steps, one can prepare and analyze protein lysate. The process involves several important steps like lysate preparation, electrophoresis, film transfer, antibody incubation, and color development. Each step is critical in ensuring the accuracy and reliability of protein analysis results.