Time-resolved fluoroimmunoassay of 5-methyl-2'-deoxycytidine.
We describe time-resolved fluoroimmunoassay of 5-methyl-2'-deoxycytidine (5MedCyd). The assay is based on the use of a highly specific antiserum raised in rabbits against BSA-conjugated 5-methylcytidine (5MeCyd). The tracer in the solid-phase time-resolved fluoroimmunoassay (TR-FIA) was antigen-selected anti-5MedCyd labeled with Europium. Thyroglobulin-linked 5MeCyd served as the solid-phase antigen. The measuring range for the fluoroimmunoassay was from less than 1 to 5000 pmol per assay of 5MedCyd. A good correlation between the results obtained with the TR-FIA and HPLC was demonstrated when the methods were applied to the measurement of methylation in human leukemic cells and other DNA samples. TR-FIA has several advantages over the more laborious techniques available so far: (i) high sensitivity, (ii) large assay ranges, (iii) rapidity and large number of simultaneous assays, (iv) simplicity, and (v) low cost provided that the laboratory has equipment for time-resolved fluorometry.
D000818:Animals; D002417:Cattle; D002851:Chromatography, High Pressure Liquid; D004247:DNA / Q000032:analysis; D003841:Deoxycytidine / Q000031:analogs & derivatives* / Q000032:analysis; D005399:Fishes; D006801:Humans; D007118:Immunoassay / Q000379:methods; D008297:Male; D013045:Species Specificity; D013050:Spectrometry, Fluorescence; D013094:Spermatozoa; D013950:Thymus Gland
D003841:Deoxycytidine; D004247:DNA; C016569:5-methyldeoxycytidine