BrdU Cell Proliferation Assay
Halogenated nucleotides such as the pyrimidine analog bromodeoxyuridine
(BrdU) can be used to label nascent DNA in viable cells. During the process
of DNA replication, BrdU can replace thymidine and incorporate into the
synthesized DNA of actively dividing cells. Since this BrdU reagent does not
cross react with endogenous DNA, cell proliferation thus can be reflected in
the extent of BrdU incorporation by determining the intensity of absorbance of
the final reaction. The BrdU cell proliferation assay is a versatile and
convenient method to quantify cell proliferation.
In the BrudU cell proliferation assay, cells are cultured with labeling medium
that contains BrdU and BrdU is incorporated into de novo-synthesized DNA
as a substitute for thymidine, which could label proliferating and daughter cells
until it is diluted out through multiple rounds of cell division. Cells proliferation
can be labeled by this method and BrdU can be detected by specific
monoclonal antibodies. There are several kinds of antibodies that can be used
to recognize the bound detection antibody. To achieve a better result, HRP
substrate TMB is added to develop color and the magnitude of the
absorbance for the developed color is proportional to the quantity of BrdU
incorporated into cells. Cell proliferation can be directly indicated by the
quantity of BrdU that is incorporated into cells. The BrdU cell proliferation
assay has been widely applied in the study of cell proliferation and this
method can also be used to assess cell proliferation in vivo.
The BrdU cell proliferation assay is a rapid, simple and high sensitive method
to assess cell proliferation. With the high resolution data of cell proliferation
acquired from this assay, a further understanding of the mechanisms of cell
proliferation can be achieved and many molecular mediators that could regulate
cell proliferation can be identified including hormones, signaling components
and transcriptional factors. Custom service is also available. This assay can be
modified to generate a more targeted result according to the purpose of the
research.
