BIOMAT Database Logo BIOMAT Database Logo

The highly abundant urinary metabolite urobilin interferes with the bicinchoninic acid assay. 2013

D L Sampson, and Y L Chng, and Z Upton, and C P Hurst, and A W Parker, and T J Parker
Tissue Repair and Regeneration Program, Brisbane, QLD 4059, Australia. Electronic address: dayle.sampson@qut.edu.au.

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 (Cu(2+)) to cuprous ions (Cu(1+)), 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.

UI MeSH Term Description Entries
D011506 Proteins Linear POLYPEPTIDES that are synthesized on RIBOSOMES and may be further modified, crosslinked, cleaved, or assembled into complex proteins with several subunits. The specific sequence of AMINO ACIDS determines the shape the polypeptide will take, during PROTEIN FOLDING, and the function of the protein. Gene Products, Protein,Gene Proteins,Protein,Protein Gene Products,Proteins, Gene
D011804 Quinolines
D012016 Reference Values The range or frequency distribution of a measurement in a population (of organisms, organs or things) that has not been selected for the presence of disease or abnormality. Normal Range,Normal Values,Reference Ranges,Normal Ranges,Normal Value,Range, Normal,Range, Reference,Ranges, Normal,Ranges, Reference,Reference Range,Reference Value,Value, Normal,Value, Reference,Values, Normal,Values, Reference
D003300 Copper A heavy metal trace element with the atomic symbol Cu, atomic number 29, and atomic weight 63.55. Copper-63,Copper 63
D006801 Humans Members of the species Homo sapiens. Homo sapiens,Man (Taxonomy),Human,Man, Modern,Modern Man
D012680 Sensitivity and Specificity Binary classification measures to assess test results. Sensitivity or recall rate is the proportion of true positives. Specificity is the probability of correctly determining the absence of a condition. (From Last, Dictionary of Epidemiology, 2d ed) Specificity,Sensitivity,Specificity and Sensitivity
D014557 Urobilin

Related Publications

D L Sampson, and Y L Chng, and Z Upton, and C P Hurst, and A W Parker, and T J Parker
Sulfo-N-hydroxysuccinimide interferes with bicinchoninic acid protein assay.
October 2011, Analytical biochemistry,
D L Sampson, and Y L Chng, and Z Upton, and C P Hurst, and A W Parker, and T J Parker
Use of the bicinchoninic acid assay in measuring urinary proteins.
March 1986, Clinical chemistry,
D L Sampson, and Y L Chng, and Z Upton, and C P Hurst, and A W Parker, and T J Parker
The bicinchoninic acid (BCA) assay for protein quantitation.
January 1994, Methods in molecular biology (Clifton, N.J.),
D L Sampson, and Y L Chng, and Z Upton, and C P Hurst, and A W Parker, and T J Parker
Interference of N-hydroxysuccinimide with bicinchoninic acid protein assay.
July 2011, Biochemical and biophysical research communications,
D L Sampson, and Y L Chng, and Z Upton, and C P Hurst, and A W Parker, and T J Parker
Bicinchoninic acid (BCA) assay in low volume.
March 2011, Analytical biochemistry,
D L Sampson, and Y L Chng, and Z Upton, and C P Hurst, and A W Parker, and T J Parker
Microplate assay of ascorbic acid in aqueous humor with bicinchoninic acid.
June 1997, Journal of ocular pharmacology and therapeutics : the official journal of the Association for Ocular Pharmacology and Therapeutics,
D L Sampson, and Y L Chng, and Z Upton, and C P Hurst, and A W Parker, and T J Parker
Drug interference in the 2,2'-bicinchoninic acid protein assay.
February 1992, Biochemical Society transactions,
D L Sampson, and Y L Chng, and Z Upton, and C P Hurst, and A W Parker, and T J Parker
Protein analysis with bicinchoninic acid.
January 1988, Annals of clinical and laboratory science,
D L Sampson, and Y L Chng, and Z Upton, and C P Hurst, and A W Parker, and T J Parker
[Urinary urobilinogen and urobilin].
August 1999, Nihon rinsho. Japanese journal of clinical medicine,
D L Sampson, and Y L Chng, and Z Upton, and C P Hurst, and A W Parker, and T J Parker
Determination of polyurethane-grafted peptide (GSGREDVGSG) using bicinchoninic acid assay.
June 2018, BioTechniques,
Copied contents to your clipboard!