SDS replacement by triton x100

Warren Gallin wgallin at gpu.srv.ualberta.ca
Tue May 3 16:18:14 EST 1994


In Article <03MAY94.16368843.0043.MUSIC at SEMOVM>, C749SCB at SEMOVM.SEMO.EDU
(Walt Lilly) wrote:
>
>A few weeks ago we asked a question about the regarding soaking
>SDS gels in triton x100 to renature proteins.  I can't believe none
>of you netters wanted to take a flyer on this.  So I thought I'd try
>again.  Here's the scoop.  We run SDS gels that contain gelatin to
>detect protease activity.  The samples are mixed with SDS loading
>buffer but not heated.  Following the run, we get no clearing of the
>gelatin unless we soak the gel in 2.5% triton X-100 for about a half
>an hour.  I'd like to know the action of the triton x-100 in causing the
>reactivation/renaturation.  Any ideas out there?

   TritonX-100 is non-denaturing.  It forms mixed micelles with SDS.  Thus,
when you soak the gel in a TritonX-100 solution the denaturing SDS is
removed and the protein can renature.  The Triton may also act to help the
renaturation by stabilizing folding intermediates that expose hydrophobic
internal regions (this is highly speculative).
   TritonX-100 has been used for renaturation of protiens on blots for
ligand probing, even after the sample was boiled, so your result fits in
with other, well established results.


Warren Gallin,
Department of Zoology, University of Alberta
wgallin at gpu.srv.ualberta.ca



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