FLAG system and salt

Matt Thomas mthomas at hsc.usf.edu
Fri Dec 1 15:21:51 EST 2000


I'm working with a complex of proteins, one of which is flag tagged.  I need
to mess with salt conditions and maybe chaotropics salts but don't realy
know how it will affect the FLAG binding.  Does anyone have any experience
with running flag columns at highish salt concentrations?  I'm thinking on
the order of 500 to 750mM KCl.  Also, how high can you go with NP-40 or
Triton before you lose binding of the flag epitope?  I'm using Sigma's Flag
resin for these experiments (a bit pricey I know, not my idea!).

Basically I want to get an idea of who "tight" the complex is and see if I
add salts who comes off the complex first before I lose binding of the flag
tagged protein to the resign.

thanks a bunch.
Matt Thomas
-- 
___________________________________________
Matthew Thomas, Ph. D
H. Lee Moffitt Cancer Center, University of South Florida
mthomas at hsc.usf.edu

"I refrain from publishing for fear that disputes and
controversies may be raised against me by ignoramuses."
   Sir Isaac Newton, correspondence to Liebniz






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