Removing all cell surface proteins/receptors - (Dec/09/2008 )
hey guys I was wondering if anyone had any ideas how to remove (either temporarily or permanently) all cell surface receptors/proteins while maintaining cell viability?
I was thinking about immersing the cells in a trypsin/EDTA mixture... any ideas?
Any help is appreciated,
thanks in advance,
I think trypsin should do the trick. EDTA is not necessary, as this does not cut proteins. An old colleague of mine used trypsin too, she used some kind of membrane dye to show that the cell membranes were still intact. Forgot the name of the dye though, so sorry, not really helping.
Any idea of showing that receptors etc have been degraded though?
Not without killing the cells . But you could try isolating the treated cells (centrifuge?) and then do a Western Blot or something similar on a standard receptor(s) compared to untreated cells. I am not completely sure though if the free proteins will remain in the cell pellet after centrifuging. Perhaps someone else could answer this?
Anyways, you could also use histochemistry to show the localization of some extracellular proteins in treated versus untreated cells. This will be slightly more work though.