i'm currently being frustrated by trying to run my protein through a semi-dry blotting system.... the extracted cell protein (i'm actually looking for UCP-2, 31kDa) is eletrophoresing very weel (as demonstrated by coomassie) but i haven't managed to get it to transfer yet.
i've tried both continuous and discontinuous buffer systems, and a range of power settings (15V for 30m/1h..... 150mA for 1h.....250mA for 1h....mA per cm2 of gel for 1 h) and none of them have caused protein to show up on ponceau s staining....
i'm using sigma's semi-dry western blotter if that helps
where am i going wrong?
Edited by flashboy, 25 March 2004 - 01:22 AM.