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Mol Biotechnol. 2013 Nov;55(3):217-26. doi: 10.1007/s12033-013-9672-6.

A defined methodology for reliable quantification of Western blot data.

Author information

  • 1Bio-Rad Laboratories (Canada) Ltd, 1329 Meyerside Dr, Mississauga, ON, L5T 1C9, Canada, sean_taylor@bio-rad.com.

Abstract

Chemiluminescent western blotting has been in common practice for over three decades, but its use as a quantitative method for measuring the relative expression of the target proteins is still debatable. This is mainly due to the various steps, techniques, reagents, and detection methods that are used to obtain the associated data. In order to have confidence in densitometric data from western blots, researchers should be able to demonstrate statistically significant fold differences in protein expression. This entails a necessary evolution of the procedures, controls, and the analysis methods. We describe a methodology to obtain reliable quantitative data from chemiluminescent western blots using standardization procedures coupled with the updated reagents and detection methods.

PMID:
23709336
[PubMed - indexed for MEDLINE]
PMCID:
PMC3840294
Free PMC Article

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