Peptide ELISA

Deef Suus News deefsuus at casema.net
Fri Feb 4 13:25:33 EST 2000


I have used KLH-conjugates to raise antibodies in rabbits. I coated the
plates with either the peptide alone or coupled to ovalbumine. Both were
dissolved in sodiumcarbonate buffer. I had no problems with the ELISA. I had
nice titers with good duplos.
Maybe your peptide doesn't dissolve in the sodium carbopnate buffer (you
might try to add some other solvent to dissolve your peptide.
I don;'t know whether that will interfere with the procedure.
Good luck

Suzanne

Tom Barr <mdp96tab at shef.ac.uk> schreef in berichtnieuws
38971108.2A37343A at shef.ac.uk...
> Hello all,
>     I've been struggling for some time now trying to measure mouse
> antibody responses to  synthetic peptides which have been  coupled to
> various carrier proteins. Does anyone out there have experience in
> peptide ELISAs who could perhaps give me some helpful advice? I've tried
> conjugating with glutaraldehyde and EDC to OVA for coating, UV treatment
> of plates, high pH buffers, poly lysine and even drying the antigen onto
> the plate... all with no success. Any advice/tips would be greatly
> appreciated...
> Thanks in advance,
>
> Tom Barr (mdp96tab at shef.ac.uk)
> Division of Molecular and Genetic Medicine,
> University of Sheffield Medical School,
> Beech Hill Road,
> Sheffield.
> S10 2RX
> Tel.: (0114) 271 2983
>






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