Improved Western Blot Assays Using BlotCycler™ Automated Wash System

Immunodetection of proteins is often performed manually and involves multiple steps that easily introduce bias and errors. We have compared the traditional manual Western blotting and a novel type of automatic Western blotting by using BlotCycler™ processor developed by Precision Biosystems, and DyLight™ fluorochrome conjugated secondary antibodies developed by Rockland Immunochemicals. The results of manual and automatic processing were comparable at RT, but automatic processing yielded consistently higher signal at 4ºC. Automatic processing using BlotCycler™ showed excellent reproducibility and can be efficiently used for optimization of primary and secondary antibodies concentration.