Quantitative Measurement of Cell Proliferation,,,Using the BrdU ELISA: A Comparison,,,Between Colorimetric and Chemiluminescent Detection
Quantitative determination of DNA synthesis in cellcultures is now a routine procedure in many laboratories.Protocols are available for various applicationsespecially in cell culture systems. The effects of growthfactors inhibition of cell division by exogenous factors
Quantitative determination of DNA synthesis in cell
cultures is now a routine procedure in many laboratories.
Protocols are available for various applications,
especially in cell culture systems. The effects of growth
factors, inhibition of cell division by exogenous factors,
stimulation/inhibition by cytokines, development of
serum-free media, influence of hormones and receptor
activity on proliferation, and selection advantages for
aneuploid cells in vitro are just a few examples where
the determination of DNA synthesis provides important
information.
In the past, radioactive tagging of newly synthesized
DNA with 3H-labeled thymidine was the most frequently
applied method, while today non-radioactive labeling
with BrdU, a thymidine analog, is finding increasing use.
Although the advantages of non-radioactive immune
techniques using BrdU or anti-BrdU are not disputed,
the scope and limitations of the available systems for
detecting BrdU are not always clearly defined. These
practical aspects have been examined in more detail in
the experiments described below. Besides enabling
microscopic detection at the cellular level, the ELISA
technique represents a method for routine screening.
Two different detection systems are available: a) the
conventional colorimetric test and b) a chemiluminescence
assay also based on the anti-BrdU technique.
One of these test methods will be suitable for use
depending on the problem and the available laboratory
facilities.
We are particularly interested in the cultivation of
human prostatic epithelial cells through several
passages. For this purpose, a special culture medium
containing exogenous growth factors and added
hormones was developed [1]. In the course of this work
we hav
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