TY - JOUR
T1 - Fractionation of Chromosomal Nonhistone Proteins Using Chromatin-Cellulose Resins
T2 - Purification of Steroid Hormone Acceptor Proteins
AU - Spelsberg, Thomas C.
AU - Stake, Eric
AU - Witzke, David
N1 - Funding Information:
The authors thank Mr. Paul Matthai, Mrs. Patti Midthun, and Mrs. Kay Rasmussen for their excellent technical help. This work was financed by grants CA 14920 and HD 9140 from the NIH and the Mayo Foundation.
PY - 1978/1/1
Y1 - 1978/1/1
N2 - 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.
AB - 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.
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U2 - 10.1016/S0091-679X(08)61151-6
DO - 10.1016/S0091-679X(08)61151-6
M3 - Article
C2 - 703618
AN - SCOPUS:0017795554
SN - 0091-679X
VL - 17
SP - 303
EP - 324
JO - Methods in cell biology
JF - Methods in cell biology
IS - C
ER -