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BrdU is an analog of thymidine readily incorporated into DNA during DNA synthesis. It provides an accurate method of monitoring proliferation and apoptosis. The Mouse Anti-BrdU antibody, clone Bu20a, is suitable for flow cytometry. The following methods were used and provide a useful guide for using anti-BrdU antibodies.
The acid treatment to unwind the DNA may affect surface immunophenotyping. Staining of cells with BrdU using DNAse I may be applicable if this is required.
Appropriate controls should be carried out for flow cytometry, consider including the following:
For all multicolor flow cytometry experiments include compensation controls and fluorescence minus one (FMO) controls, which assist with identifying gating boundaries.