cross-linking antibodyies to beads

Irit Rappley via methods%40net.bio.net (by irappley from scripps.edu)
Tue Jan 19 15:34:14 EST 2010


Thanks! I will try your suggestions.
Irit


On Jan 18, 2010, at 11:02 PM, WS wrote:

> Dear Irit,
>
> Seems to me your AB does not bind to protein G as desired, and AB
> +target binds better.
>
> You might try covalent cross-linking instead of protein G. There are
> e.g. carboxyl activated (magnetic) beads available. The coupling is
> easy: buffer exchange your AB by gel filtration into e.g. borate
> buffer, incubate with activated beads for 2hrs, quench and block, then
> wash with your target buffer.
>
> Another option is biotinylating your AB and using streptavidin coated
> beads. Procedure is similar as above.
>
> Concerning BSA as control, I'd prefer an unrelated antibody of the
> same IgG subclass instead. Should have less possible "side effects".
>
> HTH
>
> Wo
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