Hi everyone. I am now focusing on a protein which has the size of 44KD. I am using 15% SDS-PAGE for electrophoresis, nitrocellulose membrane, and Bio-Rad semi-dry transfer system. For transfer, I tried 20V for 20 min. The marker transfered well but there were still lots of proteins left on the gel which I saw in coomassie blue staining. Then the bands were wery weak on he film.
Does everyone can give me some advices for the transfer condition or tell me how to calculate the optimal voltage and minutes for transfer? Thanks