The electroimmunofixation sandwich method is described as a simple, clear and accurate technique for a reconstruction test of isoenzymes and enzyme-linked immunoglobulins. The procedure is; the isoenzymes in an enzyme specimen are separated by electrophoresis, and previously prepared enzyme-linked immunoglobulin is layered on the support media. The media are incubated at 26°C for 1h, and complexes of the isoenzyme and the enzyme-linked immunoglobulin are formed. Since these complexes are soluble type, anti-immunoglobulin antibody is layered on the media. During the second incubation at 26°C for 2h, insoluble complexes of the isoenzyme, the enzymelinked immunoglobulin and the anti-immunoglobulin antibody are formed. Then, the support media are deproteinized, and enzyme staining is performed. Free isoenzymes which are not bound with the enzyme-linked immunoglobulin are removed by the deproteinization because the enzymes are not fixed on the media. From the patterns, the enzyme-linked immunoglobulin is found to have an affinity for the isoenzymes remaining on the media. This method was applied to the GOT-linked IgG (λ), amylase-linked IgA (λ) and LDH-linked IgA (λ), and satisfactory data were resulted.
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