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., 细胞增殖测定包括一组重要的基于荧光的测试，可通过胸苷掺入评估DNA合成来监测细胞健康和细胞分裂。 溴脱氧尿苷（5-溴-2'-脱氧尿苷，BrdU）是是胸苷类似物的合成核苷。 BrdU通常用于检测活组织中的增殖细胞。 BrdU可以掺入到复制细胞的新合成的DNA中（在细胞周期的S期期间），在DNA复制期间替代胸苷。 然后可以使用BrdU特异性的抗体来检测掺入的化学物质，从而指示活跃地复制其DNA的细胞。 抗体的结合需要DNA变性，通常通过将细胞暴露于酸或热。 通常通过用缀合物抗BrdU抗体和DNA染料碘化丙啶（PI）标记以进行细胞周期分析，在流式细胞术中分析BrdU的掺入。