I have to do a western blotting for observing the expression of specific protein of interest. Cells were treated for a specific time with a specific substance and incubation time is over. Now can we keep the cells at 4 c or at -20 for some time without having any false effect on the cell line.
I agree with Thomas in my experience with aderence cells, I remove the media, wash with PBS, put on ice and scrape the cells in cold PBS, centrifuge, remove all the PBS and freeze the pellet at -80. Using a scraper with culture flask it is only a little annoying (maybe in the future it is better to use culture dishes), so you can decide to use trypsin. Someone, after washing the cells and removing PBS, freeze directly the dishes at -80.
yes i agree with the them. more specifically you should wash and then freeze them in liq N2
As an addendum to the above answer, I can add that sometimes I also freeze down whole dishes...I remove the media, wash in pbs(x3), take out all the remaining pbs and then freeze @-80 for some days (up to 1week). When I need for doing for example WB, I just add lysis buffer to the frozen dishes...two important consideration: for mRNA analysis is better not to do, and also for sensitive antigen (i.e. phosphorylation) and also be aware that it is important to freeze them dried...
if you gonna process the samples soon you can freeze at -80, don't need put in liq N2. And very importante... after take out the cells from culture wash the pellet at least two times with PBS because the serum from medium interferes in the analysis. After washing, aspirate the PBS and freeze just the pellet.
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