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Dear Michael and Grace,Thank you very much for sharing this...

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Kitson Liew May 23, 2017
protocol Protocol: Senescence Associated β-galactosidase Staining
Dear Michael and Grace,

Thank you very much for sharing this wonderful protocol.

May I know if I can keep the fixed cells in PBS at 4 C as a pause step after fixing and washing twice with PBS? If yes, would there be a limit to how long I can keep the fixed cells in su Read more Read less more less
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Kitson Liew Answered May 26, 2017

Institute for Medical Research, Kuala Lumpur, Malaysia

Thank you very much, Michael for your reply!

May I know if immunofluorescence (IF) staining can still be carried out on fixed cells previously stained for SAbeta-gal activity?

If yes, what would be the recommended procedures for (IF) after the SAbeta-gal staining?

Many thanks once again.
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Michael Eccles Author Answered May 25, 2017

Department of Pathology, University of Otago, New Zealand

We stained cells immediately after fixation. Please note that the fixation time is short, only 5min. I would suggest to process the cells as promptly as you can. If you have to, I would suggest to store fixed cells in PBS, seal it with parafilm and keep the samples at 4 degree Celsius. I would process the cells within 2-3 days, PBS salt crystals would eventually form in PBS and could affect staining/imaging. I would run a trial to compare if there is any staining positivity with the different storage period (immediate processing vs. after 2-3 days).
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