期刊
ELECTROPHORESIS
卷 28, 期 18, 页码 3163-3173出版社
WILEY
DOI: 10.1002/elps.200700002
关键词
blotting errors; high throughput; modified Western immunoblotting; protein electrophoresis; quantitative analysis
资金
- NIAAA NIH HHS [R01 AA008714, R01 AA015311, T32 AA007463, R01 AA015311-04, AA015311] Funding Source: Medline
- NIGMS NIH HHS [GM59570, R01 GM059570-06, R01 GM059570, R01 GM059570-08A1] Funding Source: Medline
The qualitative and quantitative measurements of protein abundance and protein modification states are essential in understanding their role in diverse cellular processes. Traditional Western blotting technique, though sensitive, is prone to produce substantial errors and is not readily adapted to high-throughput technologies. We propose a modified immunoblotfing procedure, which is based on simultaneous transfer of proteins from multiple gelstrips onto the same membrane, and is compatible with any conventional gel electrophoresis system. As a result, the data output per single blotting cycle can readily be increased up to ten-fold. In contrast to the traditional one protein detection per electrophoresis cycle, this procedure allows simultaneous monitoring of up to nine different proteins. In addition to maintaining the ability to detect picogram quantities of protein, the modified system substantially improves data accuracy by reducing signal errors by two-fold. Multistrip Western blotting procedure allows making statistically reliable side-by-side comparisons of different or repeated sets of data. Compared to the traditional methods, this approach provides a more economical, reproducible, and effective procedure, facilitating the generation of large amounts of high-quality quantifiable data.
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