Alternative to Trypsin-EDTA?

Warren Gallin wgallin at gpu.srv.ualberta.ca
Thu Dec 21 19:09:06 EST 2000


Depending on the cell type, I have reasonable luck using 5 mM EDTA in
PBS or TBS.  You can try leaving the plate on ice or in a 37 degree
incubator to see which gives you faster release of viable cells.
	The principle is that removing calcium ions should release any adhesion
that is integrin- or cadherin-dependent.

Warren Gallin

Benny wrote:
> 
> Dear All,
> 
> I tried to study the binding of an antibody to some cell surface protein
> molecule of a turmour cell line with FACS. However, the signals seem to be
> quite low. I suspect the level of the proteins on the cells is not very
> high. To minimise the further loss of protein during trypsinisation, I 've
> heard of one reagent that detaches cells from tissue culture plastic and
> minimises this problem. Does anyone know what it is and where I can get it
> from? Thanks.
> 
> Benny.






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