| Quantification of apolipoproteins in rat serum and in cultured rat hepatocytes by enzyme-linked immunosorbent assay. | |
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MedLine Citation:
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PMID: 3085544 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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An enzyme-linked immunosorbent assay (ELISA) has been developed to measure apolipoproteins in rat serum. Nondelipidated whole serum was heat-treated at 52 degrees C for 3 h in phosphate-buffered saline containing 0.1% Tween-20 before assay. Monospecific rabbit anti-rat apolipoprotein antibodies were added to 96-well polystyrene microtiter plates which had been coated with purified rat serum apolipoproteins or unknown samples. After incubation and washing, goat anti-rabbit serum antibodies conjugated with horseradish peroxidase were added to the plates and incubated. The bound peroxidase activity was assayed after further washing. Serum apolipoprotein concentrations were calculated by comparison against purified standards that were assayed simultaneously with the unknown samples. The intraassay coefficients of variation for apolipoprotein AI, E, and AIV (Apo AI, E, and AIV) were 2.3, 4.4, and 5.3%, and interassay coefficients of variation were 6.1, 5.5, and 7.9%, respectively. The ELISA assay is sensitive to nanogram quantities of rat serum apolipoproteins and the results agree well with those measured by densitometry. The serum concentrations of Apo AI, E, and AIV of a normal fed rat were found to be 504 +/- 8, 413 +/- 20, and 262 +/- 20 micrograms/ml, respectively. When cultured as monolayers in Waymouth's medium for 1 day, rat hepatocytes secreted Apo AI, E, and AIV at rates of 2.51, 61.8, and 48.9 ng protein/mg cell protein/h. |
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Authors:
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R C Lin |
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Publication Detail:
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Type: Journal Article; Research Support, Non-U.S. Gov't; Research Support, U.S. Gov't, Non-P.H.S. |
Journal Detail:
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Title: Analytical biochemistry Volume: 154 ISSN: 0003-2697 ISO Abbreviation: Anal. Biochem. Publication Date: 1986 Apr |
Date Detail:
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Created Date: 1986-06-16 Completed Date: 1986-06-16 Revised Date: 2006-11-15 |
Medline Journal Info:
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Nlm Unique ID: 0370535 Medline TA: Anal Biochem Country: UNITED STATES |
Other Details:
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Languages: eng Pagination: 316-26 Citation Subset: IM |
Export Citation:
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APA/MLA Format Download EndNote Download BibTex |
| MeSH Terms | |
Descriptor/Qualifier:
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Animals Apolipoprotein A-I Apolipoproteins / analysis*, blood, secretion Apolipoproteins A / analysis Apolipoproteins E / analysis Cells, Cultured Densitometry Enzyme-Linked Immunosorbent Assay* Liver / analysis*, secretion Male Rats Rats, Inbred Strains |
| Chemical | |
Reg. No./Substance:
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0/Apolipoprotein A-I; 0/Apolipoproteins; 0/Apolipoproteins A; 0/Apolipoproteins E; 0/apolipoprotein A-IV |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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