Recovering proteins from Western Blots
Berry at biochem.unp.ac.za
Thu May 5 04:38:17 EST 1994
I have a Western blot of a protein, which has been immunodetected and
stained with ABTS. I want to recover the original protein intact in order to
do an electrospray mass spec study on it. Therefore, I need to firstly strip
off the antibody (and precipitated ABTS stain?), and then release the
Has anyboby any thoughts on how I could accomplish this? Presumably an
acidic wash would remove the antibody (a la immuno-affinity chromatography),
but how will this affect the binding of protiens to the nitrocellulose
membrane (i.e. will this strip ALL protein from the blot, or, conversely,
will the immunological binding be released, but the antibody remain bound to
the membrane by non-specific binding?).
Needless to say, I have *VERY* little sample available, and so trial and
error shots are out of the question.
All contributions will be gratefully received.
Department of Biochemistry
University of Natal
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