So I found the publication attached, where they describe protocol to transfer proteins of wide range in the means of two-step transfer. First they use 1.2 mA/cm2 for 1 hour to blot the small proteins and then 3.5-7.6 mA/cm2 overnight for the big ones. However, with such high current, shouldn't the small proteins go even faster?

Also, they mention doubling concentration of the Towbin buffer, but in some ROTH technical info I found that best work buffer with low ionic strength and conductivity. Kinda contradictory...

I'm not sure whether it's relevant that I use PVDF membrane.

Article A two-step procedure for efficient electrotransfer of both h...