Chapter 11 The Isolation and Purification of Histones

Abstract

Publisher Summary This chapter describes the methods for fractionating, isolating, and characterizing different histone fraction. The histone fractions F1 and F2B are lysine-rich histones, F3 is an arginine-rich histone, and the fraction F2A is a mixture of two histones components—that is, F2A1 and F2A2. F2Al is another arginine-rich histone and F2A2 is an intermediate type having a molar ratio of lysine to arginine of about one. The chapter discusses the purification of these histone fractions. Although very useful for the purification of fractions prepared on a large scale by other methods, gel exclusion chromatography is not selective enough to separate all five histone fractions during one run. The preparation of the nucleated erythrocyte specific histone F2C is discussed. The techniques for characterization of the products obtained are: total amino acid analysis, N-terminal amino acid analysis, and polyacrylamide gel electrophoresis. The histone fractions prepared and dried are white powders easily soluble in water. They should be stored in a vacuum desiccator over silica gel at room temperature.