BrdU Cell Proliferation ELISA Kit
The BrDU Cell Proliferation ELISA Kit (ab126556) is an indirect ELISA kit for the detection of BrdU incorporation into newly synthesized DNA of actively proliferating cells. It involves the incorporation of BrdU into cells grown in microtiter plates using the cell layer as the solid phase.
During the last 2 to 24 hours of culture, BrdU is added to the wells of the microtiter plate. BrdU will be incorporated into the DNA of dividing cells. To allow the binding of the antibody to the incorporated BrdU cells, the DNA must be fixed, permeabilized, and denatured.
All of this is done in one step by treating with Fixing Solution. The anti-BrdU detector monoclonal antibody is pipetted into the wells and allowed to incubate for one hour, during which time it binds to any incorporated BrdU. Unbound antibody is washed away and horseradish peroxidase-conjugated goat anti-mouse antibody is added, which binds to the detector antibody.
Horseradish peroxidase catalyzes the conversion of the chromogenic substrate tetra-methylbenzidine (TMB) from a colorless solution to a blue (or yellow solution after the addition of the stop reagent), the intensity of which is proportional to the amount of BrdU incorporated in the cells. . The colored reaction product is quantitated using a spectrophotometer.
The resulting assay is sensitive, fast, easy to perform, and applicable to high throughput samples. In addition to assessing cell proliferation, information such as cell number, morphology, and analysis of cell antigens can be obtained from a single culture.
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