Here are some tips on how to improve the transfer step for western blotting:

  • Use a pre-stained protein ladder. This will help you to track the progress of the transfer and make sure that all of the proteins are being transferred to the membrane.
  • Use a high-quality transfer buffer. A good transfer buffer will help to ensure that the proteins are transferred efficiently and evenly.
  • Use a strong voltage. A higher voltage will increase the speed of the transfer, but it is important to use a voltage that is not too high, as this can damage the proteins.
  • Check for air bubbles. Air bubbles can prevent the proteins from being transferred to the membrane. Make sure to remove any air bubbles before starting the transfer.
  • Monitor the transfer. It is important to monitor the transfer to make sure that it is progressing as expected. If the transfer is taking too long, you may need to increase the voltage.
  • Stain the gel after transfer. This will help you to confirm that the proteins have been transferred to the membrane.

Here are some additional tips that may help to improve your western blot results:

  • Use fresh reagents. Old reagents can degrade the proteins and reduce the quality of your results.
  • Work in a clean environment. Dust and dirt can contaminate your samples and interfere with the transfer.
  • Use a consistent protocol. Reproducible results are more likely to be obtained if you use a consistent protocol for each western blot.

Once the transfer is complete it is important that all processing steps are completed in controlled and reproducible manner. The Precision Biosystems BlotCycler automates the blot processing steps leading to reproducible publication quality Western Blots