Please help me with the SDS-PAGE issue. I cannot see the protein of interest but the dirty dots all over the blots. And the dots are likely to gather around the pore of the gel holder cassette. When I reprobed with GAPDH, I can see the GAPDH blot with these dots still.
The pattern you see comes from the sponges on the membrane used for the wet transfer. Make sure your membrane is firsts soaked in methanol to completely wet, then transfer to buffer. Also, wash your sponges thoroughly before use and rinse with distilled water. Don't put too much pressure on the sandwich holder, you should not have to force it shut.
I agree with both Gregory and Era. Try washing your sponges and transfer equipment thoroughly. I would also be careful with your membrane regarding containers etc you use for washing / staining.
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