An enzyme-linked immunoadsorbent assay for rat ceruloplasmin |
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Authors: | Robert A DiSilvestro Ellen F Barber Elizabeth A David Robert J Cousins |
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Institution: | (1) Department of Foods and Nutrition, Purdue University, 47907 W. Lafayette, IN;(2) Department of Food Science and Human Nutrition, University of Florida, 32611 Gainesville, FL |
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Abstract: | A noncompetitive, colorimetric enzyme-linked immunoadsorbent assay (ELISA)† was developed for measuring rat ceruloplasmin
in serum and in medium from culture hepatocytes. The assay utilized polystyrene immobilized antibody which bound ceruloplasmin
which then bound biotinylated antibody. The biotinylated antibody-antigen complex was detected with strepavidin-alkaline phosphatase
conjugate. Standard curves for rat ceruloplasmin were constructed in the range between 10 and 50 ng/mL. Increases of 10 ng
produced an increase inA
403 of more than 0.2. With this immunoassay, serum ceruloplasmin levels were found to average 35 mg Cp/dL in control rats and
87 mg/dL in rats with experimental inflammation. Liver parenchymal cells secreted 1.6μg Cp/5×105 cells/24h. This ELISA assay for ceruloplasmin will facilitate studies on the regulation of ceruloplasmin synthesis and secretion
in both intact rats and isolated hepatocytes. |
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Keywords: | Ceruloplasmin ELISA rat serum hepatocytes inflammation |
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