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Predicate | Object |
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rdf:type | |
lifeskim:mentions | |
pubmed:issue |
3
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pubmed:dateCreated |
1981-5-26
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pubmed:abstractText |
Three immunological methods [radioimmunoassay (RIA), enzyme-linked immunosorbent assay, and radioimmunosorbent technique] were established for quantification of the potentially mutagenic O6-ethyldeoxyguanosine (O6-EtdGuo) in DNA treated with the carcinogen ethylnitrosourea in vivo or in vitro. To obtain high-affinity antibodies for specific detection of low levels of O6-EtdGuo in small amounts of DNA (cells), different schemes were applied for immunization of rabbits with the hapten O6-ethylguanosine coupled to various carrier proteins (rat serum albumin, bovine serum albumin, keyhold limpet hemocyanin). Low-dose immunization with the hapten-keyhold limpet hemocyanin conjugate resulted in antibodies with an affinity constant of 1 to 2 X 10(10) liters/mol and very low levels of cross-reactivity with normal as well as other alkylated DNA components. The RIA (the most sensitive of the three assays) detects 0.05 pmol of O6-EtdGuo at 50% inhibition of tracer (O6-ethyl[8,5'-3H]-3'-deoxyguanosine)-antibody binding. This permits quantification by RIA of O6-EtdGuo at an O6-EtdGuo:2'-deoxyguanosine molar ratio of approximately 3 X 10(-7) in a hydrolysate of 100 micrograms of ethylated DNA. By chromatographic separation of O6-EtdGuo prior to the RIA, this value can be lowered to less than 5 X 10(-8).
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pubmed:grant | |
pubmed:language |
eng
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pubmed:journal | |
pubmed:citationSubset |
IM
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pubmed:chemical |
http://linkedlifedata.com/resource/pubmed/chemical/Antibodies,
http://linkedlifedata.com/resource/pubmed/chemical/DNA,
http://linkedlifedata.com/resource/pubmed/chemical/Deoxyguanosine,
http://linkedlifedata.com/resource/pubmed/chemical/Ethylnitrosourea,
http://linkedlifedata.com/resource/pubmed/chemical/Nitrosourea Compounds
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pubmed:status |
MEDLINE
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pubmed:month |
Mar
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pubmed:issn |
0008-5472
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pubmed:author | |
pubmed:issnType |
Print
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pubmed:volume |
40
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pubmed:owner |
NLM
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pubmed:authorsComplete |
Y
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pubmed:pagination |
887-96
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pubmed:dateRevised |
2007-11-14
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pubmed:meshHeading |
pubmed-meshheading:7471102-Animals,
pubmed-meshheading:7471102-Antibodies,
pubmed-meshheading:7471102-Antibody Specificity,
pubmed-meshheading:7471102-DNA,
pubmed-meshheading:7471102-Deoxyguanosine,
pubmed-meshheading:7471102-Enzyme-Linked Immunosorbent Assay,
pubmed-meshheading:7471102-Ethylnitrosourea,
pubmed-meshheading:7471102-Nitrosourea Compounds,
pubmed-meshheading:7471102-Radioimmunoassay,
pubmed-meshheading:7471102-Rats
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pubmed:year |
1980
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pubmed:articleTitle |
Immunological quantification by high-affinity antibodies of O6-ethyldeoxyguanosine in DNA exposed to N-ethyl-N-nitrosourea.
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pubmed:publicationType |
Journal Article,
Research Support, U.S. Gov't, P.H.S.,
Research Support, Non-U.S. Gov't
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