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Methods Mol Biol. 2009;536:149-61. doi: 10.1007/978-1-59745-542-8_17.

Multistrip western blotting to increase quantitative data output.

Author information

1
Department of Pathology, Anatomy and Cell Biology, Thomas Jefferson University, JAH 233, 1020 Locust Street, Philadelphia, PA, 19107, USA. anatoly.kiyatkin@jefferson.edu

Abstract

The qualitative and quantitative measurements of protein abundance and modification states are essential in understanding their functions in diverse cellular processes. Typical western blotting, though sensitive, is prone to produce substantial errors and is not readily adapted to high-throughput technologies. Multistrip western blotting is a modified immunoblotting procedure based on simultaneous electrophoretic transfer of proteins from multiple strips of polyacrylamide gels to a single membrane sheet. In comparison with the conventional technique, Multistrip western blotting increases the data output per single blotting cycle up to tenfold, allows concurrent monitoring of up to nine different proteins from the same loading of the sample, and substantially improves the data accuracy by reducing immunoblotting-derived signal errors. This approach enables statistically reliable comparison of different or repeated sets of data, and therefore is beneficial to apply in biomedical diagnostics, systems biology, and cell signaling research.

PMID:
19378054
PMCID:
PMC2923389
DOI:
10.1007/978-1-59745-542-8_17
[Indexed for MEDLINE]
Free PMC Article

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