conventional western blot way, which is laemmil SDS PAGE, has various pros in relation to

protein experiments.

however, I managed to apply NuPAGE gel system for observing the protein experiments.

I bought NuPAGE pre-cast gel and MOPS buffer from thermo scientific and did not buy NuPAGE gel dedicated transfer buffer.

so, I carried out transfer step with basic Tris-glycine buffer with 20%MeOH at 30V constant for 1 hr.

however, protein of interest, which is predicted 150kDa size, has a tendency to decrease

an amount of blot in comparison to laemmil gel.

isn't there any changeable condition for further improving the blot?

thank you for sharing your time and reading screed.