How Do Your Western Blots Measure Up?
Publishers and funding agencies are redefining “publication-quality” data, which may change how you capture, analyze, and present quantitative Western blot data. If you want rock solid data that nobody can question, it’s critical to understand Western blot normalization.
Many strategies for Western blot normalization exist – like total protein stains, housekeeping proteins, Stain-Free™ detection, and signaling proteins. The strategy you choose depends on the context, biology, and goal of your experiment.
Accurate data measurement and analysis is the foundation of your research findings. Image file modifications using unsupported image editing and analysis software programs can compromise the integrity of your data. Ensure that you are using a software program designed to analyze the results of your Western blot experiments that is compatible with your detection system. See a list of other best practices for imaging and publishing Western blot data.
“Oddly, the most pervasive challenges to published data we see at Cell relate to loading controls. There seems to be some misalignment among scientists regarding the importance and meaning of the actin bands in a standard western blot.”
Marcus, E. Credibility and Reproducibility. Cell. 159(5): 965-966 (2014).