Protein + alkali = ?

Dr Engelbert Buxbaum via methods%40net.bio.net (by engelbert_buxbaum At hotmail.com)
Sun Feb 4 16:08:37 EST 2007


WS wrote:

> we try to detect a certain protein with a peptide antibody on
> westerns.
> 
> At first, we do not get any signal, but after treating the membrane
> (PVDF) with an alkaline (pH12-13) solution for a short time, the
> protein may be detected

Since you have an anti-peptide antibody, the antigenic site is probably
hidden before the alkali treatment and becomes unmasked by that
treatment. This could occur in two different ways: partial hydrolysis or
unfolding from breakage of intra-molecular salt-bonds.


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