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.