Direct determination of human and rabbit apolipoprotein B selectively precipitated with butanol-isopropyl ether.
A method is described for the rapid, selective, and quantitative precipitation of apolipoprotein B from isolated hypercholesterolemic rabbit and human very low density lipoproteins (VLDL), intermediate density lipoproteins (IDL), and low density lipoproteins (LDL). Lipoprotein samples are heat-treat...
Main Authors: | , |
---|---|
Format: | Article |
Language: | English |
Published: |
Elsevier
1984-12-01
|
Series: | Journal of Lipid Research |
Online Access: | http://www.sciencedirect.com/science/article/pii/S0022227520344552 |
id |
doaj-be5a68954fdd49f4b330c367484da8a6 |
---|---|
record_format |
Article |
spelling |
doaj-be5a68954fdd49f4b330c367484da8a62021-04-25T04:16:03ZengElsevierJournal of Lipid Research0022-22751984-12-01251213801386Direct determination of human and rabbit apolipoprotein B selectively precipitated with butanol-isopropyl ether.R L KleinD B ZilversmitA method is described for the rapid, selective, and quantitative precipitation of apolipoprotein B from isolated hypercholesterolemic rabbit and human very low density lipoproteins (VLDL), intermediate density lipoproteins (IDL), and low density lipoproteins (LDL). Lipoprotein samples are heat-treated at 100 degrees C in 1% SDS. The denatured apoprotein solutions are then mixed briefly with two volumes of butanol-isopropyl ether 45:55 (v/v) to precipitate the apoB. The supernatant solutions, containing the non-apoB proteins and lipids, are removed and the apoB pellet is washed once with water. To determine apoB specific activity, the apoB pellet is resolubilized in 0.5 M NaOH by heating for 30 min at 120 degrees C. The hydrolyzed apoB protein is quantitated by fluorescence of a fluorescamine derivative. The precipitation of apoB is quantitative and selective: 99.5% of rabbit 125I-labeled LDL-apoB and 97.5% of human 125I-labeled LDL-apoB is precipitated and less than 5% of 125I-labeled HDL added to unlabeled VLDL, IDL, or LDL is precipitated. Triglyceride and cholesteryl ester contamination of the apoB pellet is less than 2% of their original radioactivities.http://www.sciencedirect.com/science/article/pii/S0022227520344552 |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
R L Klein D B Zilversmit |
spellingShingle |
R L Klein D B Zilversmit Direct determination of human and rabbit apolipoprotein B selectively precipitated with butanol-isopropyl ether. Journal of Lipid Research |
author_facet |
R L Klein D B Zilversmit |
author_sort |
R L Klein |
title |
Direct determination of human and rabbit apolipoprotein B selectively precipitated with butanol-isopropyl ether. |
title_short |
Direct determination of human and rabbit apolipoprotein B selectively precipitated with butanol-isopropyl ether. |
title_full |
Direct determination of human and rabbit apolipoprotein B selectively precipitated with butanol-isopropyl ether. |
title_fullStr |
Direct determination of human and rabbit apolipoprotein B selectively precipitated with butanol-isopropyl ether. |
title_full_unstemmed |
Direct determination of human and rabbit apolipoprotein B selectively precipitated with butanol-isopropyl ether. |
title_sort |
direct determination of human and rabbit apolipoprotein b selectively precipitated with butanol-isopropyl ether. |
publisher |
Elsevier |
series |
Journal of Lipid Research |
issn |
0022-2275 |
publishDate |
1984-12-01 |
description |
A method is described for the rapid, selective, and quantitative precipitation of apolipoprotein B from isolated hypercholesterolemic rabbit and human very low density lipoproteins (VLDL), intermediate density lipoproteins (IDL), and low density lipoproteins (LDL). Lipoprotein samples are heat-treated at 100 degrees C in 1% SDS. The denatured apoprotein solutions are then mixed briefly with two volumes of butanol-isopropyl ether 45:55 (v/v) to precipitate the apoB. The supernatant solutions, containing the non-apoB proteins and lipids, are removed and the apoB pellet is washed once with water. To determine apoB specific activity, the apoB pellet is resolubilized in 0.5 M NaOH by heating for 30 min at 120 degrees C. The hydrolyzed apoB protein is quantitated by fluorescence of a fluorescamine derivative. The precipitation of apoB is quantitative and selective: 99.5% of rabbit 125I-labeled LDL-apoB and 97.5% of human 125I-labeled LDL-apoB is precipitated and less than 5% of 125I-labeled HDL added to unlabeled VLDL, IDL, or LDL is precipitated. Triglyceride and cholesteryl ester contamination of the apoB pellet is less than 2% of their original radioactivities. |
url |
http://www.sciencedirect.com/science/article/pii/S0022227520344552 |
work_keys_str_mv |
AT rlklein directdeterminationofhumanandrabbitapolipoproteinbselectivelyprecipitatedwithbutanolisopropylether AT dbzilversmit directdeterminationofhumanandrabbitapolipoproteinbselectivelyprecipitatedwithbutanolisopropylether |
_version_ |
1721510698139254784 |