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Dear all,
We usually transfer SDS PAGE to PVDF membrane at 400mA for 60 min. I need to transfer only some amount of protein from the gel to PVDF, so that I can do western blot on this membrane and use it as a guide to isolate spots from the still remaining protein in the 2D gel.
My question is how much of proteins would have transferred if I run the transfer for just 5 min or 10 min? Will I have some proteins in the PVDF membrane already? Or do I have to run may be just 2 min or longer like 15 min?
I need to have most of my proteins in the gel and only a few to be on the transferred PVDF membrane.
Thanks for any replies.
VJ.
