Comparison of total protein concentration in skeletal muscle as measured by the Bradford and Lowry assays.
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.
Seevaratnam
Rajini
R
School of Kinesiology and Health Science, York University, Toronto, ON, Canada M3J 1P3.
Patel
Barkha P
BP
Hamadeh
Mazen J
MJ
D000818:Animals; D001681:Biological Assay / Q000379:methods*; D004396:Coloring Agents; D051379:Mice; D018482:Muscle, Skeletal / Q000737:chemistry*; D011506:Proteins / Q000032:analysis*
D004396:Coloring Agents; D011506:Proteins