When we want to carry flow cytometry for cell cycle analysis or for apoptosis study as examples, we have to use Trypsin to harvest the cell without EDTA. Why do we have to use Trypsin without EDTA? What happens if we use EDTA in our Trypsin?
Hi,
as you wish to analyse apoptosis, I assume you plan to use a kit suing a DNA dye (DAPI, IP, 7-AAD) with in parallel Annexin V.
Annexin V will bind to phosphatidylserin in a Ca2+ dependent manner.
All apoptosis detection kits using annexin V provide annexin V staining buffer which contain Ca2+.
To have tried labelling cell with annexin V without Ca2+ (by mistake)...; it dosen't work.
That's why you should harvest your cells with trypsin without EDTA.
or if your cells are hard to detach, you can use trypsin with EDTA but will definitely need 2-3 washes to remove the maximum EDTA possible.
Alexis
- Badges
- Science method