A reexamination of the chelex competitive calcium binding assay.

The chelex competitive calcium binding assay has been examined as a tool for the analysis of the kinetic parameters of calcium binding substances. Scatchard analysis of calcium binding demonstrates that chelex binds calcium with apparent negative cooperativity or with more than one class of calcium binding sites, and therefore, cannot be used to provide accurate estimations of the dissociation constant or total number of binding sites on an unknown ligand. Results presented in this study indicate that the chelex assay can be effectively used for the qualitative detection of calcium binding substances in tissue extracts or biological fluids.