As regards platelets, we find that the lipophilic dyes have a greater tendency to alter function, and sometimes antigenicity, than we would like. Another suggestion is biotinylation, which seems to be remarkably benign and remarkably stable - at least over a time frame of a few days. Ken Ault >>> "Ahern, Thomas P" <Thomas.Ahern@uvm.edu> 11/21/01 22:44 PM >>> Greetings, I'm currently using CFSE to indiscriminantly stain tumor cells. I'm concerned that the reaction with free amine groups may be altering some unique tumor-specific epitopes on the membrane surface (which I'm trying to target with cominatorial peptide libraries). Is PKH more amenable to preserving the antigenic architecture of surface proteins? I understand it uses two lipophilic 'anchor' regions which insert into the bilayer, and hence may sterically hinder access to some epitopes without actually altering their structure. Are there any problems with diffusion of this dye from cell to cell? Regards, Tom. Thomas Ahern Vermont Cancer Center E-315 Given Building Burlington, VT 05405 (802) 656-2218 voice thomas.ahern@uvm.edu
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