SDS lysate impurities - streaming on gel with SDS lysates (Feb/05/2005 )
wonder if anyone can help; I get much streaming on my SDS lysates (of breast cancer cell lines) which I don't encounter on my MPER or CHAPS lysates. Could you tell me what specific steps during an SDS lysis appear to give good clean results? Do spell it out for me if you would, am quite a novice.
my SDS lysis buffer has
60 mM TRIS
100 mM DTT
dash of bromophenol blue
I use anything from 50 - 200 mcl for lysis depending on the cell pellet size. Its the subsequent steps I'm unclear about.
when i used to use sds for lysis, i have noticed problems with clearing my lysate. my problems wer partially solved when doubling the quantity of lysis buffer. Perhaps may you try too?