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.

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Abstract

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.

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ID Code: 66484
Item Type: Journal Article
Refereed: Yes
Additional URLs:
Keywords: Urobilin; Bicinchoninic acid; Protein determination; Bile pigment; Bradford; Limits of agreement analysis
DOI: 10.1016/j.ab.2013.07.027
ISSN: 0003-2697
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

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