This chapter focuses on fractionation of chromosomal nonhistone proteins using chromatin–cellulose resins: purification of steroid hormone acceptor proteins. The preparation and handling of the resin and the fractionation of the chromosomal proteins is emphasized. The chapter describes resulting purification of the nuclear “acceptors” and outlines the method for preparation of chromatin–cellulose, nucleoacidic protein (NAP)–cellulose, and DNA–cellulose resins. The quantitative removal of chromosomal proteins from the DNA requires more than high salt solutions. The treatment with UV light is essential for binding of DNA or chromatin to cellulose that is resistant to the high salt-urea treatments. The UV treated DNA–cellulose resins are more resistant to dissociation by high salt-urea conditions compared to untreated preparations. The insoluble matrices to which the DNA is attached are celluloses, agaroses, and acrylamides. The chromatin–cellulose resins can be subjected to a variety of different salt solutions, salt-urea solutions, and other types of reagents to dissociate the proteins from the DNA.
ASJC Scopus subject areas
- Cell Biology