Am Mittwoch 06 Februar 2013 17:10:35 schrieb Yuri Pompeu:
Dear All,
I am trying to probe the existence of a disulfide bond on the surface of my
protein. I have attempted Ellman´s and my results were not as clear as I
would have hoped for. I am not a sulfur/cysteine chemist and would
Perhaps you should have a look at
http://193.146.160.29/gtb/sod/usu/$UBUG/repositorio/10320076_Hawkins.pdf.
and at An introduction to methods for analyzing thiols and disulfides:
Reactions, reagents, and practical considerations. Rosa E. Hansen 1,
Jakob R. Winther Analytical Biochemistry 394
Even so, it should run somewhat differently, and probably visibly on
SDS-PAGE. Even a single phosphorylation changes mobility, and I think I
remember having seen intramolecular disulfides change mobility (anyone else
have experience on this?)
JPK
On Wed, Feb 6, 2013 at 10:33 PM, Yuri
I never done such experiments but N-Ethylmaleimide labeling and Mass-Spec could
be the solution.
Le 6 févr. 2013 à 17:10, Yuri Pompeu yuri.pom...@ufl.edu a écrit :
Dear All,
I am trying to probe the existence of a disulfide bond on the surface of my
protein.
I have attempted Ellman´s and
Hi,
to my knowledge, Ellman's reagent detects free thiols only. (Or am I
wrong?) In contrast, with the method of Thannhauser (Thannhauser [1987],
Methods Enzymol. 143, 115-9) you can determine the total amount of thiols
(free and in disulfide bonds). Any difference between the two methods
Couldn't you just run reducing/non-reducing SDS-PAGE lanes and see the
difference?
JPK
On Wed, Feb 6, 2013 at 11:10 AM, Yuri Pompeu yuri.pom...@ufl.edu wrote:
Dear All,
I am trying to probe the existence of a disulfide bond on the surface of
my protein.
I have attempted Ellman´s and my
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Dear Yuri,
If you have access to mass spec, this should be a straight forward experiment.
Find the
reference here.
