title: A defined methodology for reliable quantification of Western blot data.
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.
authors.affiliation: Bio-Rad Laboratories (Canada) Ltd, 1329 Meyerside Dr, Mississauga, ON, L5T 1C9, Canada, sean_taylor@bio-rad.com.
authors.forename: Sean C
authors.initials: SC
authors.lastname: Taylor
authors.forename: Thomas
authors.initials: T
authors.lastname: Berkelman
authors.forename: Geetha
authors.initials: G
authors.lastname: Yadav
authors.forename: Matt
authors.initials: M
authors.lastname: Hammond
mesh_terms: D000818:Animals; D015153:Blotting, Western / Q000295:instrumentation / Q000379:methods* / Q000592:standards; D003720:Densitometry; D006367:HeLa Cells; D006801:Humans; D008163:Luminescent Measurements; D011506:Proteins / Q000032:analysis*; D012015:Reference Standards; D015203:Reproducibility of Results
chemical_list: D011506:Proteins