title: Bicinchoninic acid (BCA) assay in low volume.
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.
authors.affiliation: Department of Pathology and the Life Sciences Institute, University of Michigan, Ann Arbor, MI 48109, USA.
authors.forename: Anthony
authors.initials: A
authors.lastname: Bainor
authors.forename: Lyra
authors.initials: L
authors.lastname: Chang
authors.forename: Thomas J
authors.initials: TJ
authors.lastname: McQuade
authors.forename: Brian
authors.initials: B
authors.lastname: Webb
authors.forename: Jason E
authors.initials: JE
authors.lastname: Gestwicki
mesh_terms: D003124:Colorimetry / Q000379:methods*; D004396:Coloring Agents / Q000737:chemistry; D016014:Linear Models; D011506:Proteins / Q000032:analysis*; D011804:Quinolines / Q000737:chemistry*; D012680:Sensitivity and Specificity
chemical_list: D004396:Coloring Agents; D011506:Proteins; D011804:Quinolines; C047117:bicinchoninic acid