Although many researchers believe that a successful western blot is determined by the nature of the primary antibodies, every variable within the assay can influence data quality. For reliable and reproducible quantification of western blot, automation is key. Manual processing can lead to significant variability in results between blots.
Also the importance of the washing steps should not be under-estimated. Automated processing using Precision Biosystems’ BlotCycler™ produces excellent reproducibility, in part due to the precise timing and consistency of solution changes.
Reproducibility of Western blot processing using the BlotCycler™ from Precision Biosystems. Lane 1: HeLa whole cell lysate, lane 2: recombinant human IL-2, lane 3: HeLa whole cell lysate + recombinant human IL-2. Blocking, antibody incubations and washing steps were performed using the BlotCycler™, and the fluorescent intensity between different blots was subsequently normalized using a molecular weight standard. The automated processing demonstrated excellent reproducibility.
For more on BlotCycler, visit this page: www.blotcycler.com