Biomaterial Database

Radioimmunoassay of O6-methyldeoxyguanosine in DNA of cells alkylated in vitro and in vivo. 1983

C P Wild, and G Smart, and R Saffhill, and J M Boyle

Mouse monoclonal and rabbit polyclonal antibodies have been prepared against O6-methylguanosine complexed with bovine serum albumin. In radioimmunoassay 50% inhibition of binding of [3H]O6-methyldeoxyguanosine (O6-MedG) was obtained with 0.3 pmol or 0.16 pmol unlabelled O6-MedG using monoclonal or polyclonal antibodies having affinity constants of 3.0 X 10(9) and 5.9 X 10(9) I mol-1 respectively. Cross-reactivity with normal nucleosides and other adducts was determined. Sensitivity and reproducibility were improved by chromatographic separation using an Aminex A6 column eluted with 10 mM NH4HCO3 buffer which allowed quantitation of one molecule O6-MedG per 10(7) molecules dG in 2 mg calf thymine DNA. Values for O6-MedG from 14 samples of rat liver and kidney DNA analysed by the method described and by radiochromatography on Sephadex G10 were almost identical (correlation coefficient, 0.98). A modified procedure for the purification of 5-25 micrograms DNA from 1-5 X 10(6) fibroblasts applied to polycarbonate filters was used in providing further validation of the RIA system by measuring the persistence of O6-MedG in cell lines of known repair capacity.

UI	MeSH Term	Description	Entries
D007668	Kidney	Body organ that filters blood for the secretion of URINE and that regulates ion concentrations.	Kidneys
D008099	Liver	A large lobed glandular organ in the abdomen of vertebrates that is responsible for detoxification, metabolism, synthesis and storage of various substances.	Livers
D008214	Lymphocytes	White blood cells formed in the body's lymphoid tissue. The nucleus is round or ovoid with coarse, irregularly clumped chromatin while the cytoplasm is typically pale blue with azurophilic (if any) granules. Most lymphocytes can be classified as either T or B (with subpopulations of each), or NATURAL KILLER CELLS.	Lymphoid Cells,Cell, Lymphoid,Cells, Lymphoid,Lymphocyte,Lymphoid Cell
D008770	Methylnitrosourea	A nitrosourea compound with alkylating, carcinogenic, and mutagenic properties.	Nitrosomethylurea,N-Methyl-N-nitrosourea,NSC-23909,N Methyl N nitrosourea,NSC 23909,NSC23909
D010954	Plasmacytoma	Any discrete, presumably solitary, mass of neoplastic PLASMA CELLS either in BONE MARROW or various extramedullary sites.	Plasma Cell Tumor,Plasmocytoma,Plasma Cell Tumors,Plasmacytomas,Plasmocytomas,Tumor, Plasma Cell,Tumors, Plasma Cell
D011863	Radioimmunoassay	Classic quantitative assay for detection of antigen-antibody reactions using a radioactively labeled substance (radioligand) either directly or indirectly to measure the binding of the unlabeled substance to a specific antibody or other receptor system. Non-immunogenic substances (e.g., haptens) can be measured if coupled to larger carrier proteins (e.g., bovine gamma-globulin or human serum albumin) capable of inducing antibody formation.	Radioimmunoassays
D002417	Cattle	Domesticated bovine animals of the genus Bos, usually kept on a farm or ranch and used for the production of meat or dairy products or for heavy labor.	Beef Cow,Bos grunniens,Bos indicus,Bos indicus Cattle,Bos taurus,Cow,Cow, Domestic,Dairy Cow,Holstein Cow,Indicine Cattle,Taurine Cattle,Taurus Cattle,Yak,Zebu,Beef Cows,Bos indicus Cattles,Cattle, Bos indicus,Cattle, Indicine,Cattle, Taurine,Cattle, Taurus,Cattles, Bos indicus,Cattles, Indicine,Cattles, Taurine,Cattles, Taurus,Cow, Beef,Cow, Dairy,Cow, Holstein,Cows,Dairy Cows,Domestic Cow,Domestic Cows,Indicine Cattles,Taurine Cattles,Taurus Cattles,Yaks,Zebus
D003849	Deoxyguanosine	A nucleoside consisting of the base guanine and the sugar deoxyribose.
D004247	DNA	A deoxyribonucleotide polymer that is the primary genetic material of all cells. Eukaryotic and prokaryotic organisms normally contain DNA in a double-stranded state, yet several important biological processes transiently involve single-stranded regions. DNA, which consists of a polysugar-phosphate backbone possessing projections of purines (adenine and guanine) and pyrimidines (thymine and cytosine), forms a double helix that is held together by hydrogen bonds between these purines and pyrimidines (adenine to thymine and guanine to cytosine).	DNA, Double-Stranded,Deoxyribonucleic Acid,ds-DNA,DNA, Double Stranded,Double-Stranded DNA,ds DNA
D000478	Alkylation	The covalent bonding of an alkyl group to an organic compound. It can occur by a simple addition reaction or by substitution of another functional group.	Alkylations