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Expt3 Bradford Assay- Sample Journal Article Report F09

This method utilizes beer lamberts equation of acl

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Unformatted text preview: Another method that can be used to determine the concentration of a protein is the extinction coefficient of a protein. This method utilizes Beer-Lambert’s equation of A=ΕCl, where A is absorbance, E is the extinction coefficient of the protein, C is the concentration of the protein, and l is the path length of the container or cuvette. This is more of an algebraic method that involves inserting known values and solving for the unknown, and thus offers less room for interpretation. It also has much lower sensitivity. The Bradford assay, on the other hand, although it also relies on many of the same factors, such as absorbance and concentration, is more of a graphical method that relies on the plotting of a standard curve to determine the unknown concentrations using points on the line that correspond to the absorbencies of the unknown samples. Because outliers can be thrown out, this method offers much more room for interpretation. The use of a dye also helps to make the sensitivity of this method much higher. References: 1 Bradford, M.M. (1976). A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal. Biochem. 72, 248-254. 2 Compton, S.J. and Jones, C.G. (1985). Mechanism of dye response and interference in the Bradford protein assay. Anal. Biochem. 151(2), 369-374. 3 Sigma-Aldich, Bradford Reagent Technical Bulletin http://www....
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