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rdf:type |
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lifeskim:mentions |
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pubmed:dateCreated |
1992-8-21
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pubmed:abstractText |
We have developed a method for measurement of apolipoprotein (apo) B-48 and apo B-100 in blood and subcellular fractions of rat liver based on SDS/PAGE followed by quantitative immunoblotting using 125I-Protein A. Standard curves were prepared in each assay using apo B prepared from total rat lipoproteins by extraction with tetramethylurea. Subcellular fractions (rough and smooth endoplasmic reticulum and Golgi fractions) were prepared from rat liver and separated into membrane and cisternal-content fractions. For quantification, membrane fractions were solubilized in Triton X-100, and the apo B was immunoprecipitated before separation by SDS/PAGE and immunoblotting. Content fractions were concentrated by ultrafiltration and separated by SDS/PAGE without immunoprecipitation. Quantification of apo B in subcellular fractions and detection of apo B by immunoblotting yielded consistent results. In all fractions apo B-48 was the major form, accounting for approximately three-quarters of the total apo B. By using marker enzymes as internal standards, it was calculated that all of the apo B was recovered in the endoplasmic reticulum and Golgi fractions, with approximately 80% of each form of apo B in the endoplasmic reticulum. More than 90% of the apo B of the rough- and smooth-endoplasmic-reticulum fractions was membrane-bound, whereas approx. 33 and 15% of the apo B of the cis-enriched Golgi fractions and trans-enriched Golgi fractions respectively were membrane-bound.
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pubmed:commentsCorrections |
http://linkedlifedata.com/resource/pubmed/commentcorrection/1637294-14907713,
http://linkedlifedata.com/resource/pubmed/commentcorrection/1637294-172534,
http://linkedlifedata.com/resource/pubmed/commentcorrection/1637294-177430,
http://linkedlifedata.com/resource/pubmed/commentcorrection/1637294-196662,
http://linkedlifedata.com/resource/pubmed/commentcorrection/1637294-2066680,
http://linkedlifedata.com/resource/pubmed/commentcorrection/1637294-2161829,
http://linkedlifedata.com/resource/pubmed/commentcorrection/1637294-2187869,
http://linkedlifedata.com/resource/pubmed/commentcorrection/1637294-2188646,
http://linkedlifedata.com/resource/pubmed/commentcorrection/1637294-2229075,
http://linkedlifedata.com/resource/pubmed/commentcorrection/1637294-226139,
http://linkedlifedata.com/resource/pubmed/commentcorrection/1637294-2380638,
http://linkedlifedata.com/resource/pubmed/commentcorrection/1637294-2445342,
http://linkedlifedata.com/resource/pubmed/commentcorrection/1637294-2475561,
http://linkedlifedata.com/resource/pubmed/commentcorrection/1637294-2600545,
http://linkedlifedata.com/resource/pubmed/commentcorrection/1637294-2614266,
http://linkedlifedata.com/resource/pubmed/commentcorrection/1637294-2677202,
http://linkedlifedata.com/resource/pubmed/commentcorrection/1637294-2853718,
http://linkedlifedata.com/resource/pubmed/commentcorrection/1637294-3000603,
http://linkedlifedata.com/resource/pubmed/commentcorrection/1637294-3095664,
http://linkedlifedata.com/resource/pubmed/commentcorrection/1637294-3136166,
http://linkedlifedata.com/resource/pubmed/commentcorrection/1637294-3288504,
http://linkedlifedata.com/resource/pubmed/commentcorrection/1637294-3318851,
http://linkedlifedata.com/resource/pubmed/commentcorrection/1637294-3346254,
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http://linkedlifedata.com/resource/pubmed/commentcorrection/1637294-6232923,
http://linkedlifedata.com/resource/pubmed/commentcorrection/1637294-6298249,
http://linkedlifedata.com/resource/pubmed/commentcorrection/1637294-6435623,
http://linkedlifedata.com/resource/pubmed/commentcorrection/1637294-6530586,
http://linkedlifedata.com/resource/pubmed/commentcorrection/1637294-6563038
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pubmed:language |
eng
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pubmed:journal |
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pubmed:citationSubset |
IM
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pubmed:chemical |
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pubmed:status |
MEDLINE
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pubmed:month |
Jul
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pubmed:issn |
0264-6021
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pubmed:author |
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pubmed:issnType |
Print
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pubmed:day |
1
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pubmed:volume |
285 ( Pt 1)
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pubmed:owner |
NLM
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pubmed:authorsComplete |
Y
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pubmed:pagination |
153-9
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pubmed:dateRevised |
2009-11-18
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pubmed:meshHeading |
pubmed-meshheading:1637294-Animals,
pubmed-meshheading:1637294-Apolipoprotein B-100,
pubmed-meshheading:1637294-Apolipoprotein B-48,
pubmed-meshheading:1637294-Apolipoproteins B,
pubmed-meshheading:1637294-Blotting, Western,
pubmed-meshheading:1637294-Cell Membrane,
pubmed-meshheading:1637294-Electrophoresis, Polyacrylamide Gel,
pubmed-meshheading:1637294-Endocytosis,
pubmed-meshheading:1637294-Endoplasmic Reticulum,
pubmed-meshheading:1637294-Golgi Apparatus,
pubmed-meshheading:1637294-Liver,
pubmed-meshheading:1637294-Male,
pubmed-meshheading:1637294-Microsomes, Liver,
pubmed-meshheading:1637294-Precipitin Tests,
pubmed-meshheading:1637294-Rats,
pubmed-meshheading:1637294-Rats, Inbred Strains,
pubmed-meshheading:1637294-Subcellular Fractions
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pubmed:year |
1992
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pubmed:articleTitle |
Quantification of apolipoprotein B-48 and B-100 in rat liver endoplasmic reticulum and Golgi fractions.
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pubmed:affiliation |
Department of Molecular Biology and Biotechnology, University of Sheffield, U.K.
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pubmed:publicationType |
Journal Article,
Research Support, Non-U.S. Gov't
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