Rapid apolipoprotein E phenotyping by immunofixation in agarose
Conventional determination of apolipoprotein E isomorphs comprises ultracentrifugation of 1-5 ml serum, delipidation of very low density lipoproteins (VLDL), and isoelectric focusing (IEF) in polyacrylamide gels. In order to reduce the sample volume and to avoid nonspecific protein bands, immunoblot...
| Main Authors: | , , |
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| Format: | Article |
| Language: | English |
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Elsevier
1991-05-01
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| Series: | Journal of Lipid Research |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S0022227520420401 |
| _version_ | 1818716090771963904 |
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| author | C Luley MW Baumstark H Wieland |
| author_facet | C Luley MW Baumstark H Wieland |
| author_sort | C Luley |
| collection | DOAJ |
| description | Conventional determination of apolipoprotein E isomorphs comprises ultracentrifugation of 1-5 ml serum, delipidation of very low density lipoproteins (VLDL), and isoelectric focusing (IEF) in polyacrylamide gels. In order to reduce the sample volume and to avoid nonspecific protein bands, immunoblotting was proposed. Now we describe a methodological variant that uses 25 microliters serum, replaces ultracentrifugation by precipitation of apoE-containing lipoproteins with polyethylene glycol, and delipidation by dissolution in detergent. IEF is carried out in agarose. This allows specific immunofixation of apoE-containing bands with 10 microliters antiserum per sample. This method yields apoE patterns that are specific and well resolved. Also, it offers considerable savings of time and equipment involved. |
| first_indexed | 2024-12-17T19:13:44Z |
| format | Article |
| id | doaj.art-e2eb107f9f0b4b37a519c5be2a1a816a |
| institution | Directory Open Access Journal |
| issn | 0022-2275 |
| language | English |
| last_indexed | 2024-12-17T19:13:44Z |
| publishDate | 1991-05-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Journal of Lipid Research |
| spelling | doaj.art-e2eb107f9f0b4b37a519c5be2a1a816a2022-12-21T21:35:48ZengElsevierJournal of Lipid Research0022-22751991-05-01325880883Rapid apolipoprotein E phenotyping by immunofixation in agaroseC Luley0MW Baumstark1H Wieland2Zentrallabor der Klinik, Albert-Ludwigs-Universität, Freiburg, Germany.Zentrallabor der Klinik, Albert-Ludwigs-Universität, Freiburg, Germany.Zentrallabor der Klinik, Albert-Ludwigs-Universität, Freiburg, Germany.Conventional determination of apolipoprotein E isomorphs comprises ultracentrifugation of 1-5 ml serum, delipidation of very low density lipoproteins (VLDL), and isoelectric focusing (IEF) in polyacrylamide gels. In order to reduce the sample volume and to avoid nonspecific protein bands, immunoblotting was proposed. Now we describe a methodological variant that uses 25 microliters serum, replaces ultracentrifugation by precipitation of apoE-containing lipoproteins with polyethylene glycol, and delipidation by dissolution in detergent. IEF is carried out in agarose. This allows specific immunofixation of apoE-containing bands with 10 microliters antiserum per sample. This method yields apoE patterns that are specific and well resolved. Also, it offers considerable savings of time and equipment involved.http://www.sciencedirect.com/science/article/pii/S0022227520420401 |
| spellingShingle | C Luley MW Baumstark H Wieland Rapid apolipoprotein E phenotyping by immunofixation in agarose Journal of Lipid Research |
| title | Rapid apolipoprotein E phenotyping by immunofixation in agarose |
| title_full | Rapid apolipoprotein E phenotyping by immunofixation in agarose |
| title_fullStr | Rapid apolipoprotein E phenotyping by immunofixation in agarose |
| title_full_unstemmed | Rapid apolipoprotein E phenotyping by immunofixation in agarose |
| title_short | Rapid apolipoprotein E phenotyping by immunofixation in agarose |
| title_sort | rapid apolipoprotein e phenotyping by immunofixation in agarose |
| url | http://www.sciencedirect.com/science/article/pii/S0022227520420401 |
| work_keys_str_mv | AT cluley rapidapolipoproteinephenotypingbyimmunofixationinagarose AT mwbaumstark rapidapolipoproteinephenotypingbyimmunofixationinagarose AT hwieland rapidapolipoproteinephenotypingbyimmunofixationinagarose |