期刊
ELECTROPHORESIS
卷 36, 期 17, 页码 2046-2049出版社
WILEY
DOI: 10.1002/elps.201500138
关键词
beta-Actin; Linearity range; Loading control; Western blot analysis
资金
- Natural Scientific Foundation of China [31230066]
- National Basic Research Program of China, Ministry of Science and Technology [2012CB114101]
For the purpose of data normalization in Western blot analysis, journal editors and reviewers usually require authors to reprobe the Western blot membrane with a -actin-specific antibody after detecting the target protein. In most cases, however, -actin is overloaded, which results in a failure to detect differences in protein loading. In this study, we attempted to optimize the amount of protein loaded for -actin detection to permit suitable Western blot analysis data normalization. Our data suggest that less than 2 g of total protein should be loaded when -actin is used as a loading control. We also suggest avoiding reprobing the membrane with a -actin-specific antibody.
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