detecting hydrazide groups

ANDREI POPOV ANDREI.POPOV at bbsrc.ac.uk
Thu Apr 20 14:27:23 EST 1995


Hi
>        I am trying to attach hydrazide groups to nylon membrane so that
>I can eventually attach antibody to the Hz and make an affinity membrane. I
>have got a protocol for attaching the Hz, but I want to detect the presence of
>H
>z
>before adding antibody.
>        Normally, adding Tri-nitrobenzenesulphonic acid in borate buffer
>works like a dream. Hz goes bright red or rusty colour. I have done
>this for sephadex and cuprophan.
>        But I get nothing for nylon. Either the assay works, and there is
>no nylon, or the assay doesn't work and the nylon is there...
>        Has anybody got any ideas about this ?   

You might oxydize HRPO and couple to your membrane, but...
I cant see how you will quantitate the amount of Hz...
Besides, the membrane should be blocked by some protein
to get rid of unspecific sorbtion.
Andrei




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