Acid-Urea-Triton Polyacrylamide Gel Electrophoresis of Histones
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Acid-urea polyacrylamide gels are capable of separating basic histone proteins provided they differ sufficiently in size and/or effective charge (see Chapter 16). Separation between similarly sized and charged molecules, such as the histones H2A, H2B, and the H3 forms of most organisms, can typically not be achieved. Zweidler discovered that core histones but not linker histones (see Note 1) bind the nonionic detergent Triton (1). Generally, Triton is added to an acetic acid-urea (AU) gel system to separate core histone sequence variants and histone species with overlapping AU gel patterns. This type of gel is known as an AUT or a TAU gel. To date, a single example is known where addition of Triton X-100 has allowed separation of a nonhistone primary sequence variation, the hydrophobic replacement variant of phenylalanine by leucine in fetal hemoglobin (2).
KeywordsCore Histone Electrophoresis Buffer Hamilton Microsyringe Buffer Reservoir Concentrate Ammonium Hydroxide
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