Ni-NTA biosensor chip?

Em ekhatipoREMOVE at midway.uchicagoREMOVE.edu
Sun Apr 7 22:57:07 EST 2002


NTA chips are available from BIAcore and I used them. They are about $300 a
piece, which is expensive, but probably would be worth the price if nothing
else works for you. By nothing else I mean e.g. antibodies that recognize
epitopes on your protein close to the termini or away from interacting
surfaces. Sometimes is is more beneficial to replace 6His with cisteine in
order to use disulfhydril chemistry to immobilize your protein on the chip
(however that limits redox conditions of your experiment to mostly
oxidized).

In my case I used very "sticky" nuclear protein that, once bound to the
chip, does not come off. The latter is a big disadvantage, unless one can
find conditions that would allow regeneration of the chip (otherwise your
one measurement will cost $300, and you might need up to several hundred
measurements to accomplich a relatively short study).

Emir
University of Chicago


"John" <s330477 at student.uq.edu.au> wrote in message
news:a8qphn$3to$1 at bunyip.cc.uq.edu.au...
> Hi,
>
> I am interested if anyone has used a Ni-NTA biosensor chip to immobilise a
> His-tagged protein then analysed further protein:protein interactions with
> the His-tagged protein?
>
> We have an IAsys instrument, but I don't think they have a Ni-NTA chip.  I
> could probably get access to a BIACORE if necessary.
>
> Any opinions/experiences/recommendations would be useful.
>
> Also source and approximate cost of the Ni-NTA chips used would be also
much
> appreciated.
>
> Thanks,
> John.
>
>





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