Non-reducing micro capillary electrophoresis and analytical size exclusion chromatography

Purity of purified samples was analyzed with non-reducing MCE and analytical SEC. For non-reducing MCE, 6 μl of proteins were mixed with 21 μl of sample buffer (8.4 mM Tris-HCl pH 7.0, 7.98% Glycerol, 2.38 mM EDTA, 2.8% SDS, and 2.4 mM Iodoacetamide), heated at 85°C for 10 min, and then analyzed using a Caliper LabChip GXII Touch instrument (PerkinElmer). For analytical SEC, protein samples were analyzed on an Acquity HPLC instrument (Waters) using a BEH column (200 Å, 1.7 micron, 4.7 × 300 mm) with100 mM sodium phosphate pH 6.9, 50 mM NaCl, 7.5% ethanol as the running buffer at 0.45 ml/min flow rate.