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Quantitative Measurement of Cell Proliferation,,,Using the BrdU ELISA: A Comparison,,,Between Colorimetric and Chemiluminescent Detection


Comparable results for changes in proliferation behavior, expressed as the amount of newly synthesized DNA, were also obtained with the two detection systems in comparative studies on cultivated human umbilical vein endothelial cells. In this case, the serum-free medium from PromoCell was clearly advantageous for proliferation, showing an increase of almost 50 % in comparison with the RPMI standard medium, irrespective of whether the colorimetric assay (+50 %) or the chemiluminescence assay (+49 %) was used.

Discussion

The results show that conventional colorimetric tests exhibit a limited linear range. In general, this linearity is found in the absorption range between 0.5 and 1.5. Although most readers can also detect higher absorption values (sometimes up to 3.0), linear relationships between the measured absorption values and the actual incorporation of BrdU do not exist in these ranges. Accordingly, if one wants to document an increase in proliferation or inhibition of a cell line by exogenous growth factors, there are only two possibilities:

a) To perform the measurements in a region lying within the limits of the linearity range. This neccesitates appropriate preliminary trials.

b) To use a test exhibiting a linearity range extending over several orders of magnitude.

Overall, the use of chemiluminescence test systems is advantageous since the linearity region covers around 8-4 orders of magnitude and thus also provides the possibility of detecting and documenting extreme changes of the DNA synthesis index in one and the same experiment. The use of an extensively supplemented medium for the cultivation of human prostatic carcinoma cell lines may, at first, seem to be unnecessary on the basis of the BrdU colorim
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