An improved radioimmunoassay for the quantitation of DNA methylation.
A novel radioimmunoassay of 5MedCyd is described. The assay, employing a highly specific antiserum raised in rabbits against BSA-conjugated 5MeCyd, used 5-125iodo-2'-deoxycytidine as the tracer. The measuring range for the assay was found to be 1-1000 pmol per assay of 5MedCyd. When the methods were applied to the measurement of methylation in DNA samples a good correlation between the results obtained with the radioimmunoassay and HPLC was demonstrated. The method has several advantages over the more laborious and sophisticated techniques previously available: high sensitivity, large assay range, rapidity and potential for large number of simultaneous assays, simplicity, and low cost provided that the laboratory has equipment for gamma counting.
D044503:5-Methylcytosine; D000818:Animals; D000918:Antibody Specificity; D001972:Bromodeoxycytidine / Q000031:analogs & derivatives; D002417:Cattle; D002851:Chromatography, High Pressure Liquid; D003429:Cross Reactions; D003596:Cytosine / Q000031:analogs & derivatives* / Q000032:analysis; D004247:DNA / Q000032:analysis*; D003841:Deoxycytidine / Q000031:analogs & derivatives; D005399:Fishes; D008745:Methylation; D011817:Rabbits; D011863:Radioimmunoassay
D003841:Deoxycytidine; D001972:Bromodeoxycytidine; C011288:ibacitabine; D044503:5-Methylcytosine; D003596:Cytosine; D004247:DNA