The two wavelength p M spectrophotometric method previously introduced (1) for the determination of metal ions, and shown to be valid in the presence of small molecules, has been extended to the case of media containing macromolecules such as proteins. The interaction of calcium with bovine serum albumin, using murexide as indicator, was chosen for investigation since it permitted a comparison of the binding constant thus derived with the values found in the literature and obtained by methods based on a wider variety of principles than for any other protein.
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