Journal
SCIENCE SIGNALING
Volume 8, Issue 371, Pages -Publisher
AMER ASSOC ADVANCEMENT SCIENCE
DOI: 10.1126/scisignal.2005966
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Funding
- NIH [1-R21-AI105970]
- American Cancer Society [120668-RSG-11-047-01-DMC]
- Pew Charitable Trusts [2008-000410-006]
- David and Lucile Packard Foundation [2009-34710]
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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.
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