Cell Proliferation Assay by Flow Cytometry (BrdU and PI Staining)

Cell Proliferation assays include an important set of fluorescence-based tests that can monitor cell health and cell division by evaluating DNA synthesis through thymidine incorporation. Bromodeoxyuridine (5-bromo-2'-deoxyuridine, BrdU) is a synthetic nucleoside that is an analogue of thymidine. BrdU is commonly used in the detection of proliferating cells in living tissues. BrdU can be incorporated into the newly synthesized DNA of replicating cells (during the S phase of the cell cycle), substituting for thymidine during DNA replication. Antibodies specific for BrdU can then be used to detect the incorporated chemical, thus indicating cells that were actively replicating their DNA. Binding of the antibody requires denaturation of the DNA, usually by exposing the cells to acid or heat. The incorporation of BrdU is normally analyzed in flow cytometry by labelling with a conjugate anti-BrdU antibody and DNA dyes Propidium Iodide (PI) to perform cell cycle analysis.

BrdU

Propidium Iodide

Cell cycle