An analysis of critical factors for quantitative immunoblotting

被引：163

作者：

Janes, Kevin A. ^{[1
]}

机构：

[1] Univ Virginia, Dept Biomed Engn, Charlottesville, VA 22908 USA

来源：

SCIENCE SIGNALING | 2015年 / 8卷 / 371期

关键词：

BREAST EPITHELIAL-CELLS; REGULATORY HETEROGENEITIES; SIGNALING NETWORKS; PROTEIN; ASSAY; NITROCELLULOSE; FLUORESCENCE; SEPARATION; INHIBITION; PROGRAMS;

D O I：

10.1126/scisignal.2005966

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

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.

引用

页数：11

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