Tumor generated in mouse with chemical agents. Need to study above mentioned parameters after sacrificing the mouse.
Would prefer a dye base flow cytometry assay rather than anibody based assya.
Hi Debasis
For cell cycle you can verify DNA content by Propidium Iodide using permeabilized cells.
For apoptosis you can also use PI (but without permeabilization) and Annexin V
For ROS you can use DCFDA or CellRox from Thermo Fisher
For mitochondrial membrane potential you can use JC1.
Thanks Andre but I need protocols for running flow cytometry analysis on deaggregated cell. Which may differ from general protocols used for cells from assorted cell lines.
I need details of buffer, incubation period, dye concentration and precautions for deaggregated tumor cells.
There are commercially available kits for apoptosis assay (which can be done by flow cytometry or fluorescent microscope) and cell cycle analysis.it has to be validated before you start process with samples.
Thanks Deepika,
I have gone through the details of commercially available kits. But I believe none of them are optimized for deaggregated cells. I would look forward to inputs from people who have hands on experience as literature available regarding such model is also scarce.
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