I want to kill endothelial cells that exocytose Weible Palade bodies after 15 minutes of agonist and preserve/retain non-exocytotic cells. A complement death assay against CD62P failed. Any ideas?
Hi Michael,
It's a bit complex since I don't know what kind of experiment you do. but there is a way to do negative selection...
Try this method. I used it when I isolated alveaolar type 2 cells from mouse but I did not do the negative selection of ECs
- Incubate cells with the beads and with anti-PECAM-1 Abs. since the endothelial cells express PECAM-1 constitutively, the beads with the anti-bodies, should ideally go and bind to the cells of interest, from the cell suspension.
-the endothelial cell attached to the magnetic beads can be recovered using a magnet.
- collect the rest of the supension which do not contain the ECs with a plastic pipette
Best wishes!!
Thanks Celimene
I use adherent HUVEC. CD62 transiently expressed before re-internalizing .
Ab selection will not because trypsin will interfere.
.
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