SDS-PAGE
Proteins were separated by one-dimensional SDS-PAGE using 4–15% TGX precast gels (Biorad) at 180 V for 45 min. Prior to gel loading, samples were incubated at 95°C for 3 min in loading buffer containing β-mercaptoethanol for reducing gels, or loading buffer without a reducing agent at 75°C for 10 m for non-reducing gels. Pre-stained KaleidoscopeTM protein standard, or Precision Plus Protein™ Dual Colour Standard (Biorad) were used as markers if immunotransfer was subsequently undertaken, while unstained Precision Plus Protein™ (Biorad) standard was used for gels that were to be stained by Coomassie blue.