Has anyone used an alternative tranfer buffer, for a semi-dry tranfer? Currently we are using a Thermo-Fisher branded one, but its running out and the recipe/reagents are propietary. Does anyone have an alternative protocol for this?
The most commonly used transfer buffer is Towbin's (doi:10.1073/pnas.76.9.4350, in effect SDS-PAGE running buffer with both SDS to increase the mobility of proteins and methanol to reduce it). doi:10.1006/abio.1993.1313 uses separate buffers on the membrane and gel side to resolve this conflict. In my hands, however, Dunn's buffer (10 mM NaHCO3, 3 mM Na2CO3, doi:10.1016/0003-2697(86)90207-1) is cheaper and works better.
I also find that tank blotters work better than semi-dry, probably because the Joulean heat produced by the current is distributed into a larger volume.
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