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The Bradford protein assay (also known as the Coomassie protein assay) was developed by Marion M. Bradford in 1976. [1] It is a quick and accurate [2] spectroscopic analytical procedure used to measure the concentration of protein in a solution. The reaction is dependent on the amino acid composition of the measured proteins.
The extinction law's primary application is in chemical analysis, where it underlies the Beer–Lambert law, commonly called Beer's law. Beer's law states that a beam of visible light passing through a chemical solution of fixed geometry experiences absorption proportional to the solute concentration .
The absorbance of a material that has only one absorbing species also depends on the pathlength and the concentration of the species, according to the Beer–Lambert law =, where ε is the molar absorption coefficient of that material; c is the molar concentration of those species; ℓ is the path length.
When using spectrophotometric analysis to determine the concentration of DNA or RNA, the Beer–Lambert law is used to determine unknown concentrations without the need for standard curves. In essence, the Beer Lambert Law makes it possible to relate the amount of light absorbed to the concentration of the absorbing molecule.
Several reliable methods for quantifying protein have been developed to simplify the process. These methods include Warburg–Christian method, Lowry assay, and Bradford assay (all of which rely on absorbance properties of macromolecules). Bradford assay method uses a dye to bind to protein. Most commonly, Coomassie brilliant blue G-250 dye is ...
Bradford was born October 28, 1946, in Rome, Georgia, US, and received his B.A. from Shorter College there in 1967. [1] In 1971 he married Janet Holliday. [1] [8] He obtained his Ph.D. in biochemistry from the University of Georgia in 1975, and his use of the Coomassie Brilliant Blue G-250 dye to detect proteins, which became known as the Bradford assay, was patented in 1976.
Scientists use this instrument to measure the amount of compounds in a sample. If the compound is more concentrated more light will be absorbed by the sample; within small ranges, the Beer–Lambert law holds and the absorbance between samples vary with concentration linearly. In the case of printing measurements two alternative settings are ...
It is assumed that the Beer–Lambert law applies. A = l ∑ ε c {\displaystyle A=l\sum {\varepsilon c}} where l is the optical path length, ε is a molar absorbance at unit path length and c is a concentration.