4.5. Western Blot Analysis

Spore coat proteins extracted from the purified spores of recombinant B. subtilis BV were processed by sodium dodecyl sulfate (SDS)-dithiothreitol (DTT) treatment at 65 °C for 10 min [52], and the protein solution was concentrated by freeze-drying. Concentrated proteins were fractionated on 12% denaturing polyacrylamide gels, electrotransferred onto a PVDF membrane, and used for Western blot analysis by standard procedures. Western blot filters were visualized by the enhanced chemiluminescence method as specified by the manufacturer.