CERULOPLASMIN IN HUMAN CEREBROSPINAL FLUID

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Abstract
Twenty cerebrospinal fluid samples of normal protein and cell content were investigated. After concentration by vacuum dialysis to about 1/250 of their original volume, the samples were subjected to agar gel electrophoresis on microscope slides. In order to localize ceruloplasmin on the slides 3 substrates for the oxidase activity were used: dimethyl-p-phenylene diamine, p-phenylene diamine and o-dianisidine. As the most sensitive indicator o-dianisidine was chosen for quantitative determination of ceruloplasmin. The stable red-brown color produced by oxidation of o-dianisidine was scanned at 480 nm in a spectrophotometer. In all samples investigated cerulo-plasmin was detected as a single protein fraction with the same relative mobility and oxidase activity as serum ceruloplasmin. The quantitative determination was done by comparison with an electro-phoretically pure human ceruloplasmin. Due to loss of protein the vacuum dialysis the concentration of ceruloplasmin could only be estimated roughly to 1-5 microgram copper /ml. The determination of ceruloplasmin on the slides was done with a relative standard deviation of 15%.

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