Aldrin Gomes, Associate Professor of Neurobiology at UC Davis, talks about key success factors to instill trust in western blotting data. Western blotting is a widely used technique to identify and quantify proteins in complex samples. Unfortunately, as an editor of several journals, Prof. Gomes routinely observes Western blots that are not properly conducted. The generally low quality of western blotting data has led scientists to question whether we can trust protein quantification when using western blots. Reducing errors and misinterpretations requires standardized procedure and proper controls. In this webinar, Prof. Gomes discusses the following key factors for high-confidence western blot data:
1) Quality of the primary and secondary antibodies and the blocking reagent
2) Proper target protein controls 3) Choice of total protein loading control vs. housekeeping protein loading control to normalize the target protein measurement.
Prof. Gomes also presents tips to reduce the overall time required for first-rate western blots and how to minimize the pain of going through a long procedure. The scientific community needs to continue to increase the awareness of problems associated with western blotting and introduce, improve, and standardize basic criteria for western blotting practices so that the most investigators will be successful in eliminating common problems associated with “bad” western blots.