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BrdU Staining of Cells for Cell Cycle Analysis and Apoptosis for Flow Cytometry
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.