I was wondering if this simple cytoplasmic / nuclear fractionation would work for mammalian epithelial cells. Use NP40 lysis buffer on the cell pellet for cytoplasmic fraction then pellet nuclei. Lyse nuclei with RIPA and spin down to obtain nuclear soluble fraction. Lastly, nuclear the remaining pellet can be sonicated or Mnase digested for obtaining the chromatin fraction.
Any suggestions or obvious problems that I am missing with this approach?