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Predicate | Object |
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rdf:type | |
lifeskim:mentions | |
pubmed:issue |
2
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pubmed:dateCreated |
1990-4-26
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pubmed:abstractText |
A method was developed to isolate cell surface proteins by a simple two-step procedure. Hepatocyte cell surface proteins were labeled by a cleavable biotin derivative in a covalent pulse reaction. Under the described conditions, NHS-SS-biotin proved to be an impermeant, cell surface-specific label which does not affect the impermeant, cell surface-specific label which does not affect the viability of rat hepatocytes. Biotinylated cell surface proteins could be selectively separated under non-denaturing conditions from non-biotinylated proteins and biotin-containing carboxylases by avidin affinity chromatography and sulfhydryl-mediated elution. Subsequent to alkylation of the eluted protein, individual cell surface proteins could be isolated by immunoprecipitation as shown for a selected Mr 120,000 glycoprotein gp120 of the hepatocyte plasma membrane. Using this technique, a transit time of gp120 from the endoplasmic reticulum to the cell surface of 2 h was determined. The results show that the combination of labeling with a cleavable biotin derivative, non-denaturing avidin affinity chromatography and immunoprecipitation is a useful method to isolate and study individual cell surface proteins.
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pubmed:language |
eng
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pubmed:journal | |
pubmed:citationSubset |
IM
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pubmed:chemical | |
pubmed:status |
MEDLINE
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pubmed:month |
Dec
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pubmed:issn |
0171-9335
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pubmed:author | |
pubmed:issnType |
Print
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pubmed:volume |
50
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pubmed:owner |
NLM
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pubmed:authorsComplete |
Y
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pubmed:pagination |
257-62
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pubmed:dateRevised |
2006-11-15
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pubmed:meshHeading |
pubmed-meshheading:2627937-Alkylation,
pubmed-meshheading:2627937-Animals,
pubmed-meshheading:2627937-Biological Transport,
pubmed-meshheading:2627937-Biotin,
pubmed-meshheading:2627937-Cells, Cultured,
pubmed-meshheading:2627937-Chromatography, Affinity,
pubmed-meshheading:2627937-Histocytochemistry,
pubmed-meshheading:2627937-Humans,
pubmed-meshheading:2627937-Liver,
pubmed-meshheading:2627937-Male,
pubmed-meshheading:2627937-Membrane Glycoproteins,
pubmed-meshheading:2627937-Membrane Proteins,
pubmed-meshheading:2627937-Microscopy, Electron,
pubmed-meshheading:2627937-Precipitin Tests,
pubmed-meshheading:2627937-Rats,
pubmed-meshheading:2627937-Rats, Inbred Strains,
pubmed-meshheading:2627937-Time Factors
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pubmed:year |
1989
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pubmed:articleTitle |
Selective isolation of individual cell surface proteins from tissue culture cells by a cleavable biotin label.
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pubmed:affiliation |
Institut für Klinische Chemie und Biochemie, Freie Universität, Berlin-West/Federal Republic of Germany.
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pubmed:publicationType |
Journal Article,
Research Support, Non-U.S. Gov't
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