Hi Adam, Yes we analyse MMPs and ADAMs by flow cytometry on tumour cell cultures. We use fresh cells (adherent cells detached using EDTA (0.2%, 0.2ug/ml) for up to 10 minutes at 37oC), stained with Ab (30 mins 4oC), wash, then second stage goat anti mouse or rabbit (as appropriate) F(ab')2 FITC Ig (30 mins 4oC), wash, counterstain with 10ug/ml propidium iodide (50ul in 300ul cell sample volume) to eliminate dead cells. We also fix (1%paraformaldehyde 5 mins RT), wash, and permeabilise (70% ethanol, overnight 4oC). Then stain as above to check for any cytoplasmic/inner membrane staining. I hope this helps. Best wishes, John.
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