Does anyone have a ChIP protocol that will allow me to perform IP on the nuclear fraction with chromatin, while keeping the cytoplasmic proteins so I can run a traditional IP on a Western? I would like to be able to IP the nucleus and do PCR and also IP the cytoplasm and run a Western.
Many protocols call for a cell lysis buffer to pellet the nuclei; can I just simply keep this sup for a Western? They also use 10ml of the cell lysis buffer to pellet the nuclei; can I get away with using less in order to concentrate the proteins?
Alternatively, can I perform regular ChIP up until the IP portion, but then add DNase to get rid of the chromatin? Ideally, I would like to isolate the nuclear transcription factors and perform an IP then a Western to see some co-factors.
Edit: Please delete this topic.
Edited by labstud4, 15 August 2013 - 08:48 PM.