Quantitation of 5-bromo-2-deoxyuridine incorporation into DNA: an enzyme immunoassay for the assessment of the lymphoid cell proliferative response.
As an alternative to the measurement of radiolabeled thymidine incorporated into DNA, a method is presented in which thymidine has been replaced by its analogue, 5-bromo-2-deoxyuridine (BUdR). BUdR incorporated into DNA (BUdR-DNA) is measured by a sandwich-type enzyme immunoassay using a monoclonal anti-BUdR antibody. This method allows the quantitation of 4 ng of BUdR-DNA. Comparative experiments with myeloma cells and LPS stimulated spleen B-cells have shown that this technique is at least as sensitive as the traditional counting of [3H]thymidine.
D000818:Animals; D000911:Antibodies, Monoclonal; D001973:Bromodeoxyuridine / Q000032:analysis; D004247:DNA / Q000096:biosynthesis*; D006863:Hydrogen-Ion Concentration; D007124:Immunoenzyme Techniques; D008213:Lymphocyte Activation; D008214:Lymphocytes / Q000166:cytology / Q000378:metabolism*; D051379:Mice; D009691:Nucleic Acid Denaturation; D012967:Sodium Dodecyl Sulfate; D013936:Thymidine
D000911:Antibodies, Monoclonal; D012967:Sodium Dodecyl Sulfate; D004247:DNA; D001973:Bromodeoxyuridine; D013936:Thymidine