Hi all, I am trying to work out phospho-mTOR using some muscle extracts, one problem stuck me that theoretically Bio Rad Precision Plus Protein marker doesn't appear signals in WB, however in order to have a better result, I need to run the gel (6%) till very button until 250kDa marker appear in the middle of the gel. Howver when I reveal the membrane there is always strong signal in the marker (as you see almost the whole lane) and because in my sample phopho-mTOR seems to me very low and the strong signal of marker can weaken the sample bands' signal. If I cut the marker lane and expose the samples I can see my bands, not very strong signal but I do see (picture not shown). Have you faced some similar situations like this? Thanks a lot!