High Resolution Isoelectric Focusing in a Narrow pH Interval for the Phenotyping of Vitamin D-Binding Protein (Gc-Globulin)

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Abstract
A routine method for the analysis of Vitamin D-binding protein (Gc-globulin)phenotypes was developed. The assay is based on isoelectric focusing in a narrow pH interval (4.5-5.4) using Agarose IEF (1%) or polyacrylamide (T5% C3%) gels, followed by immunofixation. When focused in this pH gradient the anodal and cathodal bands of the phenotype Gc 1-2 are separated by 16-18 mm. The fast and slow bands of the phenotype Gc 1-1 can also be easily distinguished from each other. The isoelectric focusing technique which was developed resolves the Vitamin D-binding protein phenotypes in a fast, convenient and reproducible manner. Since agarose gels are so easy to cast and process Agarose IEF is recommended as the matrix of choice.