4.5.2. Tris/Borate/EDTA Polyacrylamide Gel Electrophoresis (TBE-PAGE)

TBE-PAGE was prepared using 10× TBE buffer (Bioland) and reagents from Fluka (SDS-PAGE Kit). The 1.5 mm gels were run in a MiniProtean system (Biorad©), using 0.1 M Tris/borate, 1 mM NaEDTA, and pH 8.3 buffer. GAG (2 mg/mL) were mixed (1:5) with 2 M sucrose (Sigma) 0.02% bromophenol blue (Sigma) in 1× TBE. Gels were run at constant 60 V for 15 min and then at 150 V for 60 min at RT. After, gel was stained with 0.5% alcian blue in 2% acetic acid for 30 min; excess of staining dye was removed by rinsing the gel in 2% acetic acid for 30 min.

All the staining/washes steps were performed on an orbital shaker at room temperature.
