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Gel electrophoresis

BW Benjamin Wiseman
RN Ram Gopal Nitharwal
GW Göran Widmalm
MH Martin Högbom
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Nu-PAGE Bis–Tris gradient (4–12%) gels (Thermo Fisher Scientific) were loaded with the appropriate samples mixed with loading buffer (40 mM Tris-HCl, pH 6.8, 8% glycerol, 1% SDS, and 1 mg/mL bromophenol blue) and run at 200 V for 40 min in MES running buffer. The gels were stained with a standard 3 mg/mL Coomassie R-250 staining solution and destained with a 10% ethanol, 10% acetic acid solution. Novex Native gels (4–16%) Bis–Tris (Thermo Fisher Scientific) were loaded with the samples. Light-blue or clear buffer was used as the cathode buffer. The gels were run at 150 V for 2 h 30 min at 4 °C. NativeMark Unstained Protein Standard (Thermo Fisher Scientific) was used as a marker. The gels were stained and fixed according to the manufacturer’s protocol.

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