Resolving gel of pore size 11.25% and stacking gel of pore 5% size were prepared for separation of proteins.
80 μg of total proteins solution was mixed in equal volume of 2× Laemmli buffer [100 mM Tris buffer (pH 6.8); 4% SDS; 2% β-mercaptoethanol; 20% glycerol; 0.04% bromophenol blue] and heat denatured for 5min at 95 °C followed by incubation for 2min at 4 °C.