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.
More information about the Methods
mailing list