Mechanism
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 best used with protein concentrations of 0.01–1.0 mg/mL. and 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). The reaction mechanism is not well understood, but involves reduction of the Folin reagent and oxidation of aromatic residues (mainly tryptophan, also tyrosine). Experiments have shown that cysteine is also reactive towards to the reagent. Therefore, cysteine residues in protein probably also contribute to the absorbance seen in the Lowry Assay. The concentration of the reduced Folin reagent is measured by absorbance at 750 nm. As a result, the total concentration of protein in the sample can be deduced from the concentration of Trp and Tyr residues that reduce the Folin reagent.
The method was first proposed by Lowry in 1951. The Bicinchoninic acid assay and the Hartree–Lowry assay are subsequent modifications of the original Lowry procedure.
Read more about this topic: Lowry Protein Assay
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