In a wound healing experiment, I have multiple layers of cells in the sheet of cells. Only the cells in the lowermost layer of this cell sheet possess lamellipodium, while those on a higher plane do not. I want to perform a FACS analysis of all the cells in this cell sheet and segregate the cells that are on the lower plane and those on the higher plane to study a protein of my interest.
As the cells on the lower plane in contact with the substrate possess lamellipodia, I was thinking of marking those cells via immunofluorescence. I have come across the anti-Rac1-GTP antibody, but recent studies show that the antibody does not actually target the active form of Rac1 (Rac-GTP) but instead targets vimentin. Is there any other marker protein more expressed in cells that possess lamellipodia? Please let me know.
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