Transfer With Ease And Homemade Transfer Buffer

Protocol Overview

Run electrophoresis on standard glycine-based SDS PAGE systems or even tricine-based SDS PAGE. You can use BioRad's TGX gels or homemade gels.Soak stacks in BioRad Transfer buffer or your own homemade buffer. I have successfully used Towbin's buffer with 0.05% SDS.Place initial stack onto the transfer tray. Roll it flat.Prime your membrane in methanol (PVDF) or water (nitrocellulose) then soak in transfer buffer.Place your membrane on top of the laid down stack, and roll it flat.Take your gel and soak in transfer buffer for ~10 seconds.Place gel on top of membrane and lightly roll out any air bubbles.Place second set of stacks soaked in transfer buffer on top of the gel. Lightly roll the sandwhich.Place cathode lip on top of the tray and turn and click the lock system.If running a 7.5% gel you can run a transblot turbo 3 min protocol.If running a 10% gel you can run a transblot turbo 7 min protocol.Any gel above 10%, it is best to run a standard semi-dry (25 V for 30 min already pre-programmed) protocol. After transfer, block, probe and detect!