General description
The BCA protein assay is a simple and reliable protein
quantification method.
The BCA protein assay is based on a biuretreaction, which is
the reduction of Cu2+ toCu+ by proteins in an
alkaline solution withconcentration-dependent detection of themonovalent copper
ions. Bicinchoninic acidis a chromogenic reagent that chelates thereduced
copper, producing a purple complexwith strong absorbance at 562 nm (Smith1985,
Wiechelman 1988). This assay can beused to quantify protein concentration with
awide variety of samples and can be performedin minutes.
The Novagen® BCA Protein Assay Kit canbe used to determine
protein concentrationin the range of 20-2000 µg/ml in either astandard assay or
microassay configuration.Kit components are sufficient to complete500
standard-size reactions (50 µl proteinsample plus 1 ml reagent) or 2500
micro-scalereactions (25 µl protein sample plus 200 µlreagent). A BSA standard
(2 mg/ml) is providedfor convenient and reliable preparation ofstandard curves.
This assay is robust and can be performedin the presence of many compounds.
Somereagents, including chelating agents, strongacids or bases, and reducing
agents, interferewith the reduction and chelating reactions onwhich this assay
depends (Brown 1989). TheBCA assay is compatible with the followingNovagen
protein extraction and lysis reagents:BugBuster Protein Extraction
Reagent,PopCulture Reagent, CytoBuster ProteinExtraction Reagent, Reportasol
ExtractionBuffer, and Insect PopCulture Reagent. Optionsfor the removal or
dilution of interferingsubstances are described in the kit literature.
Other Notes
•500 mlBCA Solution
•15 ml4% Cupric Sulfate
•3 x 1 mlBSA Standard, 2 mg/ml
Smith, P.K., et al. 1985. Anal. Biochem.150,
76.
Kessler, R. J. and Fanestil, D. D. 1986. Anal. Biochem 159, 138.
Wiechelman, K., et al. 1988. Anal. Biochem.175, 231.
Brown, R., et al. 1989. Anal. Biochem.180, 136.
Legal Information
Use of the BCA protein assay is permitted for research
purposes only.
NOVAGEN is a registered trademark of Merck KGaA, Darmstadt,
Germany
Disclaimer
Toxicity: Multiple Toxicity Values, refer to MSDS (O)
