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The reagent is commonly used in the biuret protein assay, a colorimetric test used to determine protein concentration by UV/VIS spectroscopy at wavelength 540 nm. High sensitivity variants of the biuret test
The biuret test is a chemical test for proteins and polypeptides. It is based on the biuret reagent, a blue solution that turns violet upon contact with proteins, or any substance with peptide bonds. The test and reagent do not actually contain biuret; they are so named because both biuret and proteins have the same response to the test.
The traditional method for measuring total protein uses the biuret reagent, but other chemical methods such as dye-binding and refractometry are now available. The measurement is usually performed on automated analysers along with other laboratory tests. [citation needed]
The method combines the reactions of copper ions with the peptide bonds under alkaline conditions (the Biuret test) with the oxidation of aromatic protein residues. The Lowry method is based on the reaction of Cu +, produced by the oxidation of peptide bonds, with Folin–Ciocalteu reagent (a mixture of phosphotungstic acid and phosphomolybdic acid in the Folin–Ciocalteu reaction).
BCA protein assay in a 96 well plate. The bicinchoninic acid assay (BCA assay), also known as the Smith assay, after its inventor, Paul K. Smith at the Pierce Chemical Company, [1] now part of Thermo Fisher Scientific, is a biochemical assay for determining the total concentration of protein in a solution (0.5 μg/mL to 1.5 mg/mL), similar to Lowry protein assay, Bradford protein assay or ...
The Biuret assay utilizes a biuret reagent which turns purple in the presence of proteins due to the chelation of copper salts in an alkaline solution. [4] Enzyme linked immunoassays use enzyme-complexed-antibodies to detect antigens. Binding of the antibody is often inferred from the color change of reagents such as TMB.
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Biuret Test Derived Assays: Bicinchoninic acid assay (BCA assay): Detection down to 0.5 μg/mL; Lowry Protein assay: Detection in the range of 0.01–1.0 mg/mL; Fluorescamine: Quantifies proteins and peptides in solution if primary amine are present in the amino acids; Amido black: Detection in the range of 1-12 μg/mL