Measurement of protein using bicinchoninic acid

PK Smith, RI Krohn, GT Hermanson, AK Mallia… - Analytical …, 1985 - Elsevier
PK Smith, RI Krohn, GT Hermanson, AK Mallia, FH Gartner, MD Provenzano, EK Fujimoto…
Analytical biochemistry, 1985Elsevier
Bicinchoninic acid, sodium salt, is a stable, water-soluble compound capable of forming an
intense purple complex with cuprous ion (Cu1+) in an alkaline environment. This reagent
forms the basis of an analytical method capable of monitoring cuprous ion produced in the
reaction of protein with alkaline Cu2+ (biuret reaction). The color produced from this reaction
is stable and increases in a proportional fashion over a broad range of increasing protein
concentrations. When compared to the method of Lowry et al., the results reported here …
Bicinchoninic acid, sodium salt, is a stable, water-soluble compound capable of forming an intense purple complex with cuprous ion (Cu1+) in an alkaline environment. This reagent forms the basis of an analytical method capable of monitoring cuprous ion produced in the reaction of protein with alkaline Cu2+ (biuret reaction). The color produced from this reaction is stable and increases in a proportional fashion over a broad range of increasing protein concentrations. When compared to the method of Lowry et al., the results reported here demonstrate a greater tolerance of the bicinchoninate reagent toward such commonly encountered interferences as nonionic detergents and simple buffer salts. The stability of the reagent and resulting chromophore also allows for a simplified, one-step analysis and an enhanced flexibility in protocol selection. This new method maintains the high sensitivity and low protein-to-protein variation associated with the Lowry technique.
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