I'm new to the world of western blots and have had a couple attempts to run some of my cell protein. I am pouring my own 12% gels using the BioRad kits, and running them at 200V for ~30 min. With my first attempt at running my samples I had a successful transfer, however my protein ladder appeared smeared and not resolved correctly. The protein ladder I am using is PageRuler™ Plus Prestained Protein Ladder, 10 to 250 kDa.

I tried running a gel with my ladder in a few lanes and sample buffer in the remaining to ensure that it wasn't an issue with the conduction. It appeared the smearing occurred across all ladders again. A colleague suggested it may be an issue with the ladder itself and suggested I run a couple of wells with my ladders and one with one of theirs.

I ran that this morning with a newly made 10x running buffer diluted to 1x (900 mL dH20, 100 mL 10x) and upon running that I had the same issue once again with all ladders. Does anyone have any idea of what could be causing this? I am a bit at a loss.

p.s. sorry the images don't quite do it justice.