| Radioimmunoassay of apolipoprotein A-I of rat serum. | |
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MedLine Citation:
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PMID: 184835 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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A double antibody radioimmunoassay technique was developed for quantification of apolipoprotein A-I, the major apoprotein of rat high density lipoprotein. Apo A-I was labeled with 125I by the chloramine-T method. 125I-labeled apo A-I had the same electrophoretic mobility as unlabeled apo A-I and more than 80% of the 125I was precipitated by rabbit anti apo A-I antibodies. The assay is sensitive at the level of 0.5-5 ng, and has intraassay and interassay coefficients of variation of 4.5 and 6.5% respectively. The specificity of the assay was established by competitive displacement of 125I-labeled apo A-I from its antibody by apo A-I and lipoproteins containing apo A-I, but not by rat albumin and other apoproteins. Immunoreactivity of high density lipoprotein and serum was only about 35% of that of their delipidated forms when Veronal buffer was used as a diluent. Inclusion of 5 mM sodium decyl sulfate in the incubation mixture brought out reactivity equivalent to that found after delipidation. Completeness of the reaction was verified by comparison with the amount of apo A-I in chromatographic fractions of the total apoprotein of high density lipoprotein. Content (weight %, mean values +/- S.D.) of immunoassayable apo A-I was: 62.3 +/- 5.9 in high density lipoprotein; 1.7 +/- 0.3 in low density lipoprotein; 0.09 +/- 0.03 in very low density lipoprotein and 25.0 +/- 5.0 in lymp chylomicrons. Concentration in whole serum was 51.4 +/- 8.9 mg/dl and 33.6 +/- 4.1 mg/dl for female and male rats, respectively (p less than 0.002), equivalent to the sex difference in concentration of high density lipoprotein. 95% of the apo A-I in serum was in high density lipoprotein, 5% in proteins of d greater than 1.21 g/ml and less than 1% in lipoproteins of d less than 1.063 g/ml. |
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Authors:
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M Fainaru; R J Havel; T E Felker |
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Publication Detail:
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Type: Journal Article; Research Support, U.S. Gov't, P.H.S. |
Journal Detail:
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Title: Biochimica et biophysica acta Volume: 446 ISSN: 0006-3002 ISO Abbreviation: Biochim. Biophys. Acta Publication Date: 1976 Sep |
Date Detail:
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Created Date: 1976-12-30 Completed Date: 1976-12-30 Revised Date: 2006-11-15 |
Medline Journal Info:
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Nlm Unique ID: 0217513 Medline TA: Biochim Biophys Acta Country: NETHERLANDS |
Other Details:
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Languages: eng Pagination: 56-68 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 Apolipoproteins / blood* Binding Sites, Antibody Electrophoresis, Polyacrylamide Gel Female Lipoproteins, HDL / blood* Male Precipitin Tests Radioimmunoassay Rats Sodium Dodecyl Sulfate |
| Chemical | |
Reg. No./Substance:
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0/Apolipoproteins; 0/Binding Sites, Antibody; 0/Lipoproteins, HDL; 151-21-3/Sodium Dodecyl Sulfate |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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