IUBio

Protein detection problem under reducing condition

EK anybody at uchicago.edu
Wed Jun 25 08:27:44 EST 2003


""Mauro Bongrazio"" <mbon at zedat.fu-berlin.de> wrote in message
news:m19V8j4-000R1fC at Mail.ZEDAT.FU-Berlin.DE...
Hallo everybody,
I have a home-made anti-peptide antibody which detects the target
protein under non-reducing but not under reducing (beta-mercapto)
conditions. Do you know why? I thought the contrary could be the
case.Thanks
Mauro

---

Obviously, you are unfolding the antibody and thus disrupting  the
recognition surface. This is very common. Now you know that your antibody
has a surface, rather than a linear sequence, that recognizes your peptide.
BME reduces disulfide bonds that  in your case contribute to correct folding
of the Ab.
-Emir





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