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.

SUBMITTER: Janes KA 

PROVIDER: S-EPMC4401487 | biostudies-literature | 2015 Apr

REPOSITORIES: biostudies-literature