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Antibody titers

Dekkerc DEKKERC at CENTOCOR.COM
Fri Jan 28 07:10:34 EST 2000


>
>antigen: immunisation peptide coupled to ovalbumin (immunisation is done
>using peptide coupled to KLH)
>immunisation scheme:
>December 15th: retreived blood (preserum)
>December 15th: 1st injection in Freunds complete adjuvant
>January 12th: retrieved first blood (serum I)
>January 12th: 1st booster injection in Freunds incompelte adjuvant
>January 20th: retreived 2nd blood (srum II)
>I trested all three samples. The serum I and II are the same height.
>Preseurm is very low.
>Coating is done at pH 9.6
>Second antibody is HRP.
>Reaction only takes place for 5 min. otherwise it is too long.


Sounds like you have an aspecific bond that enhances the signal. Do you also
have high "blanc" values?
It is a few years ago since I designed  some ELISA's, but I remember
problems with aspecific binding very well...
Maybe it is obvious, but do you block your ELISA-plates after coating?
Sometimes the second antibody will bind on the plate, and your signal goes
up dramatically.

Cees.






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