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Fig. 2 | BMC Biochemistry

Fig. 2

From: Identification of replication-dependent and replication-independent linker histone complexes: Tpr specifically promotes replication-dependent linker histone stability

Fig. 2

Purification of H1.1, H1.2 and H1x Complexes. a–c Soluble whole cell extracts of U2OS cells expressing 6 × His - tagged H1.1 (a), H1.2 (b) or H1x (c) were purified on a Mono Q column. Top panel: 1 ml fractions were collected, resolved by SDS-PAGE gel, and immunoblotted with anti-His antibodies. Bottom panel: chromatogram showing chromatography conditions used. d–f Peaks containing 6 × His – tagged H1.1 (d), H1.2 (e) or H1x (f) from Mono Q column were pooled and applied to a nickel column in loading buffer containing 20 mM imidazole, washed with 50 mM imidazole and then eluted with 500 mM imidazole. Top panel: 1 ml fractions were collected, resolved by SDS-PAGE gel, and visualized by immunoblotting with anti-His antibodies. Bottom panel: chromatogram showing chromatography conditions used

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