Sounds good! We also use 20% methanol and 0.02% SDS so maybe that’ll do the trick. Always happy to help!

We have both in the lab, but I haven’t tried the HMW stuff with the fancy fast transfer (although it says that it should be able to handle it). 2 hours in the cold room 400mA. You’re welcome to take a couple gels if you want to try them to rule out if it’s tris-glycine or just those specific gels

I have 4-12% tris-glycine - in Titia’s lab we never bought the 4-20% tris glycine ones for some reason but maybe this is why? I have no idea why the 4-20 specifically would have a problem

I haven’t had any problem transferring a 350kDa protein with the 4-12% tris-glycine gels for 2 hours at 400mA - have you tried 4-12% gels? I have some if you want to try it