Bradford assay kit is a common method used in protein quantitation that involves the binding of protein to Staining Solution (Coomassie Brilliant Blue G-250). The simple procedure is adaptable to nearly any volume scale, including test tubes and microplates. Binding of proteins to Staining Solution under acidic environment of the reagent results in spectral shift from protonated reddish/brown form of dye (absorbance maximum at 465 nm) to the unprotonated blue form of dye (absorbance maximum at 610 nm).The difference between the two forms of the dye is greatest at 595 nm, which is the optimal wavelength to measure the blue color from the Coomassie dye-protein complex. This method is useful for determining protein concentration from 50~1,000 μg/ml.
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