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Predicate | Object |
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rdf:type | |
lifeskim:mentions | |
pubmed:issue |
5
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pubmed:dateCreated |
1989-6-8
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pubmed:abstractText |
A hybridoma (K1A8) secreting a high affinity antibody to imidazole ring-opened 7-methylguanine (N5-methyl-N5-formyl-2,5,6-triamino-4-hydroxypyrimidine) was obtained from spleen cells of a mouse immunized with a conjugate of keyhole-limpet hemocyanin and imidazole ring-opened 7-methylguanylic acid (iro-7mGMP). The antibody recognizes the iro-7-methylguanine (iro-7mG) determinant in the BSA-iro-7mGMP conjugate, in chemically methylated, denatured DNA, and in the ring-opened 7-methylguanosine, 7-methyldeoxyguanosine and 7mGMP haptens. In the competitive ELISA of DNA-iro-7mG, 50% inhibition (I50) was observed at 4 fmol determinant per well (8 x 10(-11) M) using BSA-iro-7mGMP as the immobilized antigen. The lower limit of 7-methylguanine (7mG) detection in DNA is determined by the binding of unmodified DNA per se to the antibody. The intrinsic reaction of DNA with antibody is low; in the competitive ELISA I50 was obtained with 330 micrograms calf thymus DNA per 50 microliters well, equivalent to 4 nmol iro-7mG per mol nucleotide. The 7mG content of calf thymus DNA is 7 nmol per mol nucleotide (approximately 20 amol per micrograms DNA). The limit of detection of 7mG by competitive ELISA is quoted provisionally as 7 nmol iro-7mG per mol nucleotide, where 30% inhibition of antibody binding is obtained in the presence of 105 micrograms DNA per 50 microliters well. Nuclear DNAs of tissue culture cells treated with 0, 0.01 and 0.1 mM N-methyl-N'-nitro-N-nitrosoguanidine contained 0.18, 31 and 320 mumol, respectively, of 7-methylguanine adducts per mol of nucleotides. This report indicates that the K1A8 antibody will serve to quantify DNA alkylation in human populations exposed to low levels of methylating carcinogens.
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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/7-methylguanine,
http://linkedlifedata.com/resource/pubmed/chemical/Alkylating Agents,
http://linkedlifedata.com/resource/pubmed/chemical/Antibodies, Monoclonal,
http://linkedlifedata.com/resource/pubmed/chemical/Antigen-Antibody Complex,
http://linkedlifedata.com/resource/pubmed/chemical/DNA,
http://linkedlifedata.com/resource/pubmed/chemical/Epitopes,
http://linkedlifedata.com/resource/pubmed/chemical/Guanine,
http://linkedlifedata.com/resource/pubmed/chemical/Haptens,
http://linkedlifedata.com/resource/pubmed/chemical/Methylnitronitrosoguanidine,
http://linkedlifedata.com/resource/pubmed/chemical/N(5)-methyl-N(5)-formyl-2,5,6-triami...,
http://linkedlifedata.com/resource/pubmed/chemical/Pyrimidines,
http://linkedlifedata.com/resource/pubmed/chemical/Sulfuric Acid Esters,
http://linkedlifedata.com/resource/pubmed/chemical/dimethyl sulfate
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pubmed:status |
MEDLINE
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pubmed:month |
May
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pubmed:issn |
0143-3334
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pubmed:author | |
pubmed:issnType |
Print
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pubmed:volume |
10
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pubmed:owner |
NLM
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pubmed:authorsComplete |
Y
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pubmed:pagination |
927-31
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pubmed:dateRevised |
2007-11-14
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pubmed:meshHeading |
pubmed-meshheading:2468429-Alkylating Agents,
pubmed-meshheading:2468429-Alkylation,
pubmed-meshheading:2468429-Animals,
pubmed-meshheading:2468429-Antibodies, Monoclonal,
pubmed-meshheading:2468429-Antigen-Antibody Complex,
pubmed-meshheading:2468429-Cells, Cultured,
pubmed-meshheading:2468429-DNA,
pubmed-meshheading:2468429-Enzyme-Linked Immunosorbent Assay,
pubmed-meshheading:2468429-Epitopes,
pubmed-meshheading:2468429-Guanine,
pubmed-meshheading:2468429-Haptens,
pubmed-meshheading:2468429-Hybridomas,
pubmed-meshheading:2468429-Lymphocytes,
pubmed-meshheading:2468429-Methylnitronitrosoguanidine,
pubmed-meshheading:2468429-Mice,
pubmed-meshheading:2468429-Pyrimidines,
pubmed-meshheading:2468429-Sulfuric Acid Esters
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pubmed:year |
1989
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pubmed:articleTitle |
High avidity monoclonal antibody to imidazole ring-opened 7-methylguanine.
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pubmed:affiliation |
Institute for Cell Analysis, University of Miami School of Medicine, FL 33101.
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pubmed:publicationType |
Journal Article,
Research Support, U.S. Gov't, P.H.S.
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