Protocol Online logo
Top : New Forum Archives (2009-): : SDS-PAGE and Western Blotting

Transfer failure - (Aug/14/2017 )

I just had a transfer fail in the sense that only the top band of the ladder transferred to the membrane (PVDF preactivated with methanol) and one single band of protein according to ponceau.  Meanwhile, I can see the pink of the lower ladder band on the whatman paper going the wrong way?!?!?  I am absolutely certain the transfer was electrophoresed in the proper orientation and according to my normal protocol.  But it looks like the top part of the gel transferred the correct direction and the bottom half the opposite direction?  I'm so confused and hoping someone has an answer.  Thank you!


Sounds like either you had partial transfer and then loss of power (and recovery?) or there is something wrong with the buffer such that it is missing the ions necessary for the current to be maintained properly. Diffusion can also cause proteins to be transferred out of the gel onto filterpaper and membrane at the same time.