semi-dry transfer - semi-dry transfer flop (Dec/07/2004 )

Hi,

I'm using BioRad's semi-dry transfer apparatus for transfering to PVDF membranes. In a hurry to catch a seminar, I accidently reversed the order of my gel and membrane.

Unexpectedly, after I discovered my mistake at the end of the transfer, I noticed that the membrane was well stained on both sides with the prestained ladder. There was no staining on the filter paper below the gel.

Is there any way that the protein was still transfered to the membrane? I decided to go ahead and block the membrane over night and stain the gel with commassie blue.

Thoughts...ideas...

Thanks,

John
ps I'm kind of novice with western blots so don't laugh too
hard at my ignorance

Hi,
if you also swapped over the anode/cathode wires (plugged into the wrong side, or up in the powerpoint) your blot would work as normal. That's the only thing I can think of at the moment.