I have done tens of subcellular fractionation of human cells. So what sort of cells are we looking at and what level of purity of fractions are we talking of. What sort of fractions do you want from the cells? Are they human cells, are they suspension cell cultures or monolayers or tissue samples? Try giving all the information you need before putting up a question like that. It should help to get help fast.
[Only registered users see links. ] ("Jayakumar, R") wrote in message news:<97101976F8A044468CA74FE11883B90E07496361@VIS TA.roswellpark.org>...
I wanted isolate separately cytoplasmic and nuclear proteins from
cells. These are human primary effusion lymphomas cells, which are
suspension cell cultures and latently infected by HHV8 (KSHV).The
specific problem is that my cytoplasmic extract is always contaminated
by proteins from nuclei fraction.(has been checked by cytosolic and
nuclear markers, by Western)
I tried many, many different options i.e hypotonic buffers, various
detergent and many variants of salt concetration....