title: An analysis of critical factors for quantitative immunoblotting.
abstract: Immunoblotting (also known as Western blotting) combined with digital image analysis can be a reliable method for analyzing the abundance of proteins and protein modifications, but not every immunoblot-analysis combination produces an accurate result. I illustrate how sample preparation, protocol implementation, detection scheme, and normalization approach profoundly affect the quantitative performance of immunoblotting. This study implemented diagnostic experiments that assess an immunoblot-analysis workflow for accuracy and precision. The results showed that ignoring such diagnostics can lead to pseudoquantitative immunoblot data that markedly overestimate or underestimate true differences in protein abundance.
authors: Department of Biomedical Engineering, University of Virginia, Charlottesville, VA 22908, USA. E-mail: kjanes@virginia.edu.
authors: Kevin A
authors: KA
authors: Janes
mesh_terms: D015153:Blotting, Western / Q000379:methods*; D045744:Cell Line, Tumor; D006801:Humans