How to analyse freshly isolated peripheral blood mono nuclear cells simultaneously for CD34 expression and the cell cycle phase they are in. The options I have is to use propidium iodide or hoechst for the cell cycle analysis. I would like to know which of these is a better option to combine with cell surface staining for CD34 and analysis by flow cytometry. Please provide a protocol if any.
You can use BrdU combined with 7-AAD staining to check the cell cycle and the choose a CD34 antibody that doesn't lie in the 7-AAD or BrdU channel.
Check this link for detailed protocol
http://www.bdbiosciences.com/documents/BD_FACSVerse_CellCycleAnalysis_AppNote.pdf
There is an alternative which does not require cell fixation and permeabilisation. Use DyeCycle cell permeant DNA stains after the surface staining.
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