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Separation of histone variants by RP-HPLC
Summary
Type: HPLCApplication #: 108550
Detection:
Conditions: The lyophilized proteins were dissolved in water containing 200 mM 2-mercaptoethanol and samples of 150 μg were injected onto the column. The histone sample was chromatographed within 100 min at a constant flow of 1.0 ml/min with a multi-step acetonitrile gradient starting at 40% B ((A) water containing 0.1% (v/v) TFA and 10% (v/v) ethylene glycol monomethyl ether (EGME), (B) 70% (v/v) acetonitile, 0.1% (v/v) TFA and 10% (v/v) EGME). The concentration of solvent B was increased linearly in the following order: from 40 to 61% (45 min), from 61 to 64% (35 min), from 64 to 74% (2 min), maintained at 74% for 9 min, and from 74 to 100% (10 min).
Authors: Sarg B
Source: GIT Special Chromatography 15 (1995) 50 - 54
Details
Legend: Fig. 1: Separation of whole histones by RP-HPLC. 150 µg protein samples isolated from butyrate-treated murine leukaemic cells were injected on to a NUCLEOSIL® 300-5 C4 column. Conditions see above,