Hi, I´m fixating MC3T3 cells in cold formaldehyde and then store samples in PBS. Later I want to stain these cells for ALP. Do you know how long time you can wait with the staining? Will ALP activity decrease with time? Do I have to stain immediately after fixation?
I´m a student quite new to cell biology, so any help is appreciated!
Erik
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bob1
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Thelymitra pulchella
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Posted 21 April 2010 - 04:44 PM
Formaldehyde fixes the proteins so that they are stable. The fixation will degrade over time in PBS, so that your cells will only stay fixed for a week or two. For long term storage, dehydrate your cells progressively in 25%, 50%, 75%, 100% methanol and store at -20 deg C. To use after this , rehydrate the cells using the reverse sequence of methanol.
