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rdf:type |
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lifeskim:mentions |
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pubmed:issue |
1
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pubmed:dateCreated |
1982-9-17
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pubmed:abstractText |
1. A variety of detergents were used to solubilize 5'-nucleotidase from rat liver plasma membranes. 2. The zwitterionic detergent Sulphobetaine 14 gave optimal solubilization by the criteria of release into a high-speed-centrifugation supernatant and the formation of the smallest and least polydisperse active enzyme observed on polyacrylamide-gel electrophoresis. 3. The Sulphobetaine 14-solubilized enzyme from rat liver was purified by using the conventional techniques of ion-exchange chromatography and gel filtration, or by an immunoaffinity step with a monoclonal antibody immunoadsorbent. 4. 5'-Nucleotidase was purified at least 12 000-fold relative to liver homogenate by the immunoaffinity purification scheme and had a specific activity in the range 285-340 mumol/min per mg of protein. The yield was in the range 9-16%. 5. The purified enzyme shows a major polypeptide band of apparent Mr 70 000 on sodium dodecyl sulphate/polyacrylamide-gel electrophoresis and a minor band of apparent Mr 38 000. 6. A rational approach to the general problem of the purification of minor intrinsic membrane proteins is discussed, with the use of polyacrylamide-gel electrophoresis to determine the most appropriate detergent and monoclonal antibodies in subsequent immunoaffinity purification.
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pubmed:commentsCorrections |
http://linkedlifedata.com/resource/pubmed/commentcorrection/6285894-1252510,
http://linkedlifedata.com/resource/pubmed/commentcorrection/6285894-14228777,
http://linkedlifedata.com/resource/pubmed/commentcorrection/6285894-14907713,
http://linkedlifedata.com/resource/pubmed/commentcorrection/6285894-409338,
http://linkedlifedata.com/resource/pubmed/commentcorrection/6285894-4321546,
http://linkedlifedata.com/resource/pubmed/commentcorrection/6285894-4326772,
http://linkedlifedata.com/resource/pubmed/commentcorrection/6285894-4326970,
http://linkedlifedata.com/resource/pubmed/commentcorrection/6285894-435255,
http://linkedlifedata.com/resource/pubmed/commentcorrection/6285894-4364131,
http://linkedlifedata.com/resource/pubmed/commentcorrection/6285894-4444531,
http://linkedlifedata.com/resource/pubmed/commentcorrection/6285894-4721620,
http://linkedlifedata.com/resource/pubmed/commentcorrection/6285894-479147,
http://linkedlifedata.com/resource/pubmed/commentcorrection/6285894-495956,
http://linkedlifedata.com/resource/pubmed/commentcorrection/6285894-5432063,
http://linkedlifedata.com/resource/pubmed/commentcorrection/6285894-5644126,
http://linkedlifedata.com/resource/pubmed/commentcorrection/6285894-5659650,
http://linkedlifedata.com/resource/pubmed/commentcorrection/6285894-6036837,
http://linkedlifedata.com/resource/pubmed/commentcorrection/6285894-6160155,
http://linkedlifedata.com/resource/pubmed/commentcorrection/6285894-6245642,
http://linkedlifedata.com/resource/pubmed/commentcorrection/6285894-6249264,
http://linkedlifedata.com/resource/pubmed/commentcorrection/6285894-6254034,
http://linkedlifedata.com/resource/pubmed/commentcorrection/6285894-6257695,
http://linkedlifedata.com/resource/pubmed/commentcorrection/6285894-6258567,
http://linkedlifedata.com/resource/pubmed/commentcorrection/6285894-6260546,
http://linkedlifedata.com/resource/pubmed/commentcorrection/6285894-6268444,
http://linkedlifedata.com/resource/pubmed/commentcorrection/6285894-6271180,
http://linkedlifedata.com/resource/pubmed/commentcorrection/6285894-6280685,
http://linkedlifedata.com/resource/pubmed/commentcorrection/6285894-637870,
http://linkedlifedata.com/resource/pubmed/commentcorrection/6285894-6444593,
http://linkedlifedata.com/resource/pubmed/commentcorrection/6285894-655420,
http://linkedlifedata.com/resource/pubmed/commentcorrection/6285894-677454,
http://linkedlifedata.com/resource/pubmed/commentcorrection/6285894-6935651,
http://linkedlifedata.com/resource/pubmed/commentcorrection/6285894-6966279,
http://linkedlifedata.com/resource/pubmed/commentcorrection/6285894-6970337,
http://linkedlifedata.com/resource/pubmed/commentcorrection/6285894-708065,
http://linkedlifedata.com/resource/pubmed/commentcorrection/6285894-7215667,
http://linkedlifedata.com/resource/pubmed/commentcorrection/6285894-737168,
http://linkedlifedata.com/resource/pubmed/commentcorrection/6285894-7569,
http://linkedlifedata.com/resource/pubmed/commentcorrection/6285894-851441,
http://linkedlifedata.com/resource/pubmed/commentcorrection/6285894-9017
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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 |
Apr
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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 |
203
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pubmed:owner |
NLM
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pubmed:authorsComplete |
Y
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pubmed:pagination |
245-51
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pubmed:dateRevised |
2010-9-13
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pubmed:meshHeading |
pubmed-meshheading:6285894-5'-Nucleotidase,
pubmed-meshheading:6285894-Animals,
pubmed-meshheading:6285894-Antibodies, Monoclonal,
pubmed-meshheading:6285894-Betaine,
pubmed-meshheading:6285894-Cell Membrane,
pubmed-meshheading:6285894-Centrifugation,
pubmed-meshheading:6285894-Detergents,
pubmed-meshheading:6285894-Electrophoresis, Polyacrylamide Gel,
pubmed-meshheading:6285894-Immunosorbents,
pubmed-meshheading:6285894-Liver,
pubmed-meshheading:6285894-Nucleotidases,
pubmed-meshheading:6285894-Rats,
pubmed-meshheading:6285894-Solubility
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pubmed:year |
1982
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pubmed:articleTitle |
Solubilization and purification of rat liver 5'-nucleotidase by use of a zwitterionic detergent and a monoclonal-antibody immunoadsorbent.
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pubmed:publicationType |
Journal Article,
Research Support, Non-U.S. Gov't
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