Abstract
Publisher Summary This chapter describes the methods for the fractionation of nonhistone chromosomal proteins utilizing hydroxyapatite chromatography. The hydroxyapatite procedure is a simple one-column procedure that not only separates the nonhistone proteins from the histones in high yields, but also provides a preliminary fractionation of these proteins. The chapter presents a fractionation of chromatin, using a column of hydroxyapatite. At near–neutral pH, the application of salt–urea dissociated chromatin results in the basic histones being unretained and the nonhistone proteins (and RNA) being adsorbed by the column. By using appropriate concentrations of phosphate buffer in the eluting medium, the nonhistone proteins can be eluted while the DNA is still retained by the hydroxyapatite. Electrophoretic analyses show that the three subfractions of the nonhistone proteins consist of different polypeptide species and, other analyses indicate that different biological activities can be separated by this procedure.
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