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Immunofluorescence staining of cells in combination with PI staining of cells for cell cycle analysis
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This method provides a general procedure for DNA staining for cell cycle analysis using propidium iodide when you need to stain for other intracellular antigens. 70% ethanol is optimal for DNA so if you do not need to stain for other antigens we recommend Protocol FC12 “Propidium iodide staining of cells for cell cycle analysis”. This method will allow you to stain for DNA and cellular antigens, however these are guidelines only and the incubation times may need to be adjusted for different cell types and different antibodies.
Appropriate controls should always be carried out, for flow cytometry the following should be considered for inclusion;
For all multi-color flow cytometry experiments it is advisable to include compensation controls and Fluorescence Minus One (FMO) controls, which assist with identifying gating boundaries.