I want to use FAC to study integrins on the cell surface and I've heard some people claim that Trypsin can induce integrin degradation. I'm just wondering if anyone has seen any significant differences in their results when using EDTA or Trypsin to detach their adherent cells from the plate? And if you use EDTA, which in theory don't degrade integrins then would you see a better effect?
Any insights is appreciated.
Thank you
hi
its simple . you can test two methods by same condition (antibody concentration & number of cells)and compare your result together finally . it looks good(0.02% EDTA / PBS ).
good luck
I could test yes but it just takes a lot of effort to do and I'm not even sure its worth it. That's why Im asking on here to see who's already done it and can let me know.
Anh,
Just a simple alternative to consider: grow your cells in 96- or 48-well plates and do a simple colorimetric or fluorescence-based "cell ELISA" to measure the levels of integrin(s). Very straight-forward approach and no cell detachment is required.
Best
Gedas
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