2.4. High-Resolution Clear Native (hrCN) Polyacrylamide Gel Electrophoresis (PAGE)

The hrCN-PAGE protocol was adapted from Lemaire et al. [30]. Glycerol (20% v/v final) was added to samples and 0.001% w/v Ponceau S was used as a protein migration marker. The electrophoresis cathode buffer contained a buffer mixture of 50 mM Tricine; 150 mM Bis-Tris pH 7 supplemented with 0.05% w/v sodium deoxycholate; 0.01% w/v dodecyl maltoside. The anode buffer contained 150 mM Bis-Tris buffer, pH 7. The NativeMark™ unstained protein standard from Thermo Fisher Scientific (Darmstadt, Germany) was used as a ladder. hrCN-PAGE were carried out using an 8 to 15% linear polyacrylamide gradient, gels were run with a constant 20 mA current using a PowerPac^TM Basic Power Supply (Bio-Rad). After electrophoresis, the protein bands were stained with Instant Blue^TM (Expedeon, Heidelberg, Germany).