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DMP cross-linking protocol
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Twenty-four hours after transfection, HEK-293 cells were washed twice in cold PBS and lysed on ice with lysis buffer (50 mM boric acid, 100 mM K acetate, 2 mM MgCl2, 1 mM EGTA and 1% Triton X-100; pH 8.5). One-half of the proteins was incubated with 5 mM DMP (dimethyl pimelimidate) for 1 h, while the other half was maintained for use as controls. The reaction was stopped by adding 1 ml of 0.5 M (pH 6.8). Protein samples (50 μg) were boiled in Laemmli SDS loading buffer and separated on 7% SDS-PAGE gels.
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