Re: PKH dyes

From: Kenneth Ault (aultk@mmc.org)
Date: Sat Nov 24 2001 - 07:57:56 EST


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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