The highly abundant urinary metabolite urobilin interferes with the bicinchoninic acid assay
Sampson, Dayle L., Chng, Yee Lin, Upton, Zee, Hurst, Cameron P., Parker, Anthony W., & Parker, Tony J. (2013) The highly abundant urinary metabolite urobilin interferes with the bicinchoninic acid assay. Analytical Biochemistry, 442(1), pp. 110-117.
Estimation of total protein concentration is an essential step in any protein- or peptide-centric analysis pipeline. This study demonstrates that urobilin, a breakdown product of heme and a major constituent of urine, interferes considerably with the bicinchoninic acid (BCA) assay. This interference is probably due to the propensity of urobilin to reduce cupric ions (Cu2+) to cuprous ions (Cu1+), thus mimicking the reduction of copper by proteins, which the assay was designed to do. In addition, it is demonstrated that the Bradford assay is more resistant to the influence of urobilin and other small molecules. As such, urobilin has a strong confounding effect on the estimate of total protein concentrations obtained by BCA assay and thus this assay should not be used for urinary protein quantification. It is recommended that the Bradford assay be used instead.
Impact and interest:
Citation counts are sourced monthly from and citation databases.
These databases contain citations from different subsets of available publications and different time periods and thus the citation count from each is usually different. Some works are not in either database and no count is displayed. Scopus includes citations from articles published in 1996 onwards, and Web of Science® generally from 1980 onwards.
Citations counts from theindexing service can be viewed at the linked Google Scholar™ search.
|Item Type:||Journal Article|
|Keywords:||Urobilin; Bicinchoninic acid; Protein determination; Bile pigment; Bradford; Limits of agreement analysis|
|Divisions:||Current > Schools > School of Biomedical Sciences
Current > QUT Faculties and Divisions > Faculty of Health
Current > Institutes > Institute of Health and Biomedical Innovation
|Deposited On:||22 Jan 2014 01:32|
|Last Modified:||15 Jun 2016 22:08|
Repository Staff Only: item control page