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. 2009 May 13;37(11):e81. doi: 10.1093/nar/gkp273

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© 2009 The Author(s)

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.0/uk/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

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Figure 3. — (A) Flow chart of histone purification through SP, after H₂SO₄ extraction from whole cells. (B and C) 15% SDS–PAGE analysis of single-step SP-purification of whole core histones (B) and fractionation of H2A/H2B from H3/H4 (C), respectively. (B) Following washes with 0.5 M NaCl (lanes 4–6) and with 0.6 M NaCl (lanes 7–13), histones were eluted at 2 M NaCl and further fractionated by HClO₄-precipitation into core histones (lanes 14–16) and linker histones (lanes 17–19). (C) Following the 0.6 M NaCl wash [as in (B)], H2A/H2B were separated from H3/H4 by stepwise elution at 0.8 M NaCl (lanes 2–5), followed by H3/H4 elution at 2 M NaCl (lanes 6–9). Potential problems with TCA precipitation of diluted column fractions can be overcome by another SP run (Note 4).