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Anal Biochem. 2011 Mar 15;410(2):310-2. doi: 10.1016/j.ab.2010.11.015. Epub 2010 Nov 13.

Bicinchoninic acid (BCA) assay in low volume.

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  • 1Department of Pathology and the Life Sciences Institute, University of Michigan, Ann Arbor, MI 48109, USA.

Abstract

The BCA assay is a colorimetric method for estimating protein concentration. In 96-well plates, the relationship between protein content and absorbance is nearly linear over a wide range; however, performance is reduced in lower volume. To overcome this limitation, we performed the BCA assays in opaque, white 384-well plates. These plates emit fluorescence between 450-600 nm when excited at 430 nm; thus, their fluorescence is quenched by the BCA chromophore (λ(max) 562 nm). This arrangement allowed accurate determination of protein content using only 2 μL of sample. Moreover, soluble flourescein could replace the white plates, creating a homogenous format.

PMID:
21078286
DOI:
10.1016/j.ab.2010.11.015
[PubMed - indexed for MEDLINE]
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