Can you couple avidin to an affinity chromatography matrix?
klenchin at macc.wisc.edu
Wed Nov 15 17:49:25 EST 1995
In article <email-1511951624400001 at dell.sbc.man.ac.uk>,
email at domain.com (Your Name) wrote:
>In article <Pine.A18.104.22.1681017095749.29554A-100000 at tigger.cc.uic.edu>,
>"Samuel C. Blackman" <blackman at tigger.cc.uic.edu> wrote:
>> We have an interesting idea for affinity chromatography, but it would
>> be dependant on being able to couple avidin to an affinity chromatography
>> matrix (i.e., Bio-Rad Affi-Gel 10 or 15) via a free NH2- group. Has anyone
>> tried this, or know of a refernce pointing to a method?
>Ive used affigel and other similar CBr matrices to construct affinity
>matricies and i wouldnt anticipate any problem with avidin. The pI of
>avidin is about 10 incase you need to adjust your coupling buffer. The
>only question is why bother? immobilised avidin is commercially available
>on a range of matrices already.
That's right. Wash your column with 0.5-1 M salt as avidin has considerable
nonspesific binding. Streptavidin is much better but expensive.
IMO the cheapest quality source is Molecular Probes, but everyone else sells it.
In case you want to do it yourself and save money, I'd recommend using Neutralite
(Neutra) avidin (that's deglycos. avidin with small peptide removed (pI 6.3),
essentialy the same as streptavidin but much cheaper).
The sources are Fisher (we get 40% off from it), Pierce, Sigma, Mol. Probes, etc.
/ /\ Dima Klenchin
/ / \
/ / /\ \ klenchin at macc.wisc.edu
/ / /\ \ \ tel. (608)262-4380
/ /_/__\ \ \ FAX (608)262-4570
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