Hi, I culture HACAT mammalian immortalized cells. I am interested to check the protein changes by western blot and cell cycle analysis by flow cytometry. Can i store the harvested cell pellets in -80 freezer without any storing reagent before performing western or flow cytometry?
Thank you
They should be OK for Western blotting, but I don't think they would be OK for flow cytometry because the cell membranes would be damaged by ice crystals.
For the western blot, it should be fine. You can harvest the whole protein from the cell pellet after freeze in -80 freezer.
For the cell cycle analysis, no.
As alternative, you can fix the cell in 4% formaldehyde, wash, and resuspend in PBS + 2% FBS, and keep the cell in fridge for overnight;
For the long term storage, you can cryopreserve the cells in 10% DMSO+90% FBS in-80 freezer.
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