Separation of 5'-ribonucleoside monophosphates by ion-pair reverse-phase high-performance liquid chromatography |
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| Authors: | T F Walseth G Graff M C Moos N D Goldberg |
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| Affiliation: | 1. Biophysical Endocrinology Section, Endocrinology and Reproduction Research Branch, National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, Maryland 20205 USA;2. Naval Research Laboratory, Chemistry Division, Code 6120, Washington, D. C. 20375 USA |
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| Abstract: | We have developed computer programs for characterization of ligand-binding systems in terms of continuous affinity distributions of arbitrary shape based on a numerical finite difference method. This method provides an excellent initial estimate of the affinity distribution, which can be further refined by means of nonlinear least-squares curve fitting. The method has been extensively tested for several cases including receptor heterogeneity, cooperativity, and for several examples of experimental design (e.g., ligand concentrations), and various levels of random and systematic experimental errors. The results provide a guide to experimental design, and indicate limits to the resolution obtained by ligand-binding studies, irrespective of the method of analysis. |
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