Gel Filtration Protein Molecular Weight

Standard molecular weight proteins from the high molecular weight Gel Filtration Calibration Kit (GE Healthcare) were dissolved with the running buffer (50 mM phosphate, 150 mM NaCl, pH 7.0) as described in the manual. Briefly, for standard proteins, 0.5 ml of each standard protein was loaded onto the HiLoad 26/600 Superdex 200 pg column (GE Healthcare) with a recommended flow rate of 2.6 ml/min in the AKTA purifier L25. Blue Dextran 2000 was used to detect the void volume. UV peaks of each protein were used to calculate the elution volume (24 (link)). Pfu Rnl and the fusion protein were loaded separately on the column and elution volumes were recorded for molecular weight comparison (25 (link)).