developed using the ECL techniquePerformed under reducing conditions.Exposure time : 10 seconds HeLa cells were incubated at 37&degC for 6h with vehicle control (0 &microM) and different concentrations of sodium butyrate (ab120948). Increased expression of histone H3 (acetyl K27) (ab4729) in HeLa cells correlates with an increase in sodium butyrate concentration, as described in literature.Whole cell lysates were prepared with RIPA buffer (containing protease inhibitors and sodium orthovanadate), 2.5&microg of each were loaded on the gel and the WB was run under reducing conditions. After transfer the membrane was blocked for an hour using 5% BSA before being incubated with ab4927 at 1 &microg/ml and ab8227 at 1 &microg/ml overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP (ab97051) at 1/10000 dilution and visualised using ECL development solution.

developed using the ECL techniquePerformed under reducing conditions.Exposure time : 10 seconds HeLa cells were incubated at 37°C for 6h with vehicle control (0 µM) and different concentrations of sodium butyrate (ab120948). Increased expression of histone H3 (acetyl K27) (ab4729) in HeLa cells correlates with an increase in sodium butyrate concentration, as described in literature.Whole cell lysates were prepared with RIPA buffer (containing protease inhibitors and sodium orthovanadate), 2.5µg of each were loaded on the gel and the WB was run under reducing conditions. After transfer the membrane was blocked for an hour using 5% BSA before being incubated with ab4927 at 1 µg/ml and ab8227 at 1 µg/ml overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP (ab97051) at 1/10000 dilution and visualised using ECL development solution.


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