I am trying to identify a large multi-protein complex which includes my protein of interest. I am trying to use non-denaturing gel electrophoresis (NativePAGE) to keep this complex intact, before proceeding to transfer and western blotting as well as staining of the gel. The ultimate idea is to end up with a silver stained band I can excise and analyze via mass spec. As we have little experience of this method, I was wondering if anyone had any recommendations regarding the optimal amount of protein I should load per lane and running time/ voltage. I tried using 50 ug and only obtained very faint bands. I then tried concentrating my samples and excluding all proteins