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Journal of Biochemistry Advance Access originally published online on March 6, 2009
Journal of Biochemistry 2009 145(6):791-797; doi:10.1093/jb/mvp037
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© The Authors 2009. Published by Oxford University Press on behalf of the Japanese Biochemical Society. All rights reserved

Comparison of Total Protein Concentration in Skeletal Muscle as Measured by the Bradford and Lowry Assays

Rajini Seevaratnam1,2, Barkha P. Patel1,2 and Mazen J. Hamadeh1,2,3,*

1School of Kinesiology and Health Science, York University, Toronto, ON, Canada M3J 1P3; 2Muscle Health Research Centre, York University, Toronto ON, Canada M3J 1P3; and 3Department of Pediatrics, McMaster University, Hamilton, ON, Canada L8N 3Z5

*To whom correspondence should be addressed. Tel: +1-416-736-2100 ext. 33552, Fax: +1-416-736-5774, E-mail: hamadeh{at}yorku.ca

Received January 28, 2009; Accepted February 23, 2009


  Abstract

The Lowry and Bradford assays are the most commonly used methods of total protein quantification, yet vary in several aspects. To date, no comparisons have been made in skeletal muscle. We compared total protein concentrations of mouse red and white gastrocnemius, reagent stability, protein stability and range of linearity using both assays. The Lowry averaged protein concentrations 15% higher than the Bradford with a moderate correlation (r = 0.36, P = 0.01). However, Bland–Altman analysis revealed considerable bias (15.8 ± 29.7%). Both Lowry reagents and its protein–reagent interactions were less stable over time than the Bradford. The linear range of concentration was smaller for the Lowry (0.05–0.50 mg/ml) than the Bradford (0–2.0 mg/ml). We conclude that the Bradford and Lowry measures of total protein concentration in skeletal muscle are not interchangeable. The Bradford and Lowry assays have various strengths and weaknesses in terms of substance interference and protein size. However, the Bradford provides greater reagent stability, protein–reagent stability and range of linearity, and requires less time to analyse compared to the Lowry assay.

Key Words: Lowry assay, Bradford assay, total protein concentration, skeletal muscle, Spectrophotometry

Abbreviations: BCA, bicinchoninic acid; BSA, bovine serum albumin; DTT, dithiothreitol; EDTA, ethylenedinitrilotetraacetic acid; KCl, potassium chloride; wt/vol, weight per volume


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