Binding and elution of Abs from NC bound proteins

Ed Rybicki ED at micro.uct.ac.za
Fri Aug 6 03:09:42 EST 1993


> I would be most interested in other people's experiences with this technique,
> especially regarding the buffer used for elution.

I have always used a pH 2.9 buffer, 0.1M glycine/HCl, usually with 0.1M
NaCl added.  You have to neutralise fast, but if you have pre-titrated an
amount of weak base, is no problem!  Always worked well in our hands, and
takes pretty much all (ie: >70%) of Ab off first wash.  Good Ab, too -
work in westerns and ELISA.  Can also re-use blot several times for re-
adsorption and re-elution, so doesn't mess up the adsorbed protein too
much either.

  ____________________________________________________________________
 | Ed Rybicki, PhD             |       "Lord, won't you buy me        |
 | (ed at micro.uct.ac.za)        |                                      |
 | Dept Microbiology           |         A Mer-ce-des Benz..."        |
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