I already seeded and treated the cells in 6 well plate: 2 wells treated and four wells control (2 for positive control stain and 2 for negative control stain). How to collect and store these cells as the machine of flow not ready yet?
Thanks in advance for your help.
Hi Heba,
For cell cycle assay (i.e. Propidium Iodide, PI assay), you can fix the cells in 80% Ethanol and store them at -20 degrees for a relatively long period, then you can proceed with your PI staining and readings on flow cytometry.
As for apoptosis studies, it actually depends on the assay you want to use... If Annexin V staining, you will need living cells to stain them directly with labeled Annexin. if you're going for TUNEL assay, you can fix your cells in 4% Formaldehyde and store them at +4 degrees. So it all goes back to the assay you're choosing.
Best,
Patrick
Hi Heba,
For cell cycle assay (i.e. Propidium Iodide, PI assay), you can fix the cells in 80% Ethanol and store them at -20 degrees for a relatively long period, then you can proceed with your PI staining and readings on flow cytometry.
As for apoptosis studies, it actually depends on the assay you want to use... If Annexin V staining, you will need living cells to stain them directly with labeled Annexin. if you're going for TUNEL assay, you can fix your cells in 4% Formaldehyde and store them at +4 degrees. So it all goes back to the assay you're choosing.
Best,
Patrick
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