Technical noteProtein quantification by chemiluminescent Western blotting: Elimination of the antibody factor by dilution series and calibration curve
Section snippets
Acknowledgements
We thank C. Blanc and R.K. Daher for their critical reading of the manuscript. This study was supported by the Canadian Institutes of Health Research Grant MOP-7088 and the Canada Research Chair in Stress Signal Transduction.
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A systematic approach to quantitative Western blot analysis
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2015, Neurobiology of AgingCitation Excerpt :The expression data yielded lower HZ-to-WT ratios (0.32–0.41) than did the activity data (0.49–0.51), which as mentioned previously, has also been observed by others. This discrepancy is likely because of the nonlinearity of integrated ECL signals in Western blotting, in which lower loadings of a given protein are sometimes underestimated (Charette et al., 2010). Using the same antibody used in the present study, we have previously found a similar nonlinearity when loading different amounts of pure recombinant PIMT for Western blotting.
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