Vol. 41, No. 1-2, 1997
Evaluation of the bromodeoxyuridine incorporation assay for the measurement of cell proliferation and DNA synthesis
STEC, J., REICHERT, M.
Veterinary National Research Institute
Department of Biochemistry
24-100 Pulawy
Poland
Summary
In this paper will report on the evaluation of an alternative method for the measurement of radiolabelled thymidine incorporation into DNA in which [3H]-thymidine has been replaced by its analogue 5-bromo-2-deoxyuridine (BrdUrd). The method was applied to the measurement of DNA synthesis in the mitogen induced proliferation of sheep lymphocytes. The immunoenzymatic level of bromodeoxyuridine uptake was compared with the level of [3H]-thymidine incorporation. By the method of standard linear regression it was indicated that the correlation between BrdUrd incorporation and [3H]-thymidine uptake was very high (r = 0.91, p < 10-5). The bromodeoxyuridine-ELISA assay is simple to perform, reproducable and almost as sensitive as the [3H]-thymidine test. A primary advantage of the the BrdUrd method is the fact that radioactive isotopes are not required, therefore, there are no problems with the handling and disposal of radioactive waste and the toxic scintilation fluid.
Key words: cell proliferation; DNA synthesis; sheep lymphocytes; bromodeoxyuridine; [3H]-thymidine
References
Folia Veterinaria / 1997