I'm running a 5% stacking and 10% separating gel. Gel is run at 100V for about 2 hours. Transfered to PVDF membrane at 100V overnight wet with ice block (can this be it?). Using a HRP staining system
Another problem I'm having is on the same gel but probed with a different antibody. The protein I'm interested in should just be one band at 76kDa (bottom visible horizontal lane of second gel) but I'm getting a whole bunch of staining on the top. I never used to get this. Could it be because I'm using a new lot of the same antibody from the same manufacturer? Is it non-specific staining or is my protein just getting separately improperly?
Edited by Nezura, 10 February 2011 - 07:44 PM.