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Antibody titers

spotsnall katzday at tamu.edu
Wed Jan 26 10:09:58 EST 2000


In article <388df976$0$17235 at reader3.casema.net>, deefsuus at casema.net says...
>
>Hi,
>
>I have immunised a rabbit and I was wonderign what kind of titers you can
>expect. I tested the serum and I had to dilute the serum 1:100.000 at least
>to get a reasonable ELISA-curve (even at 1:1000000 there is till a good
>signal). When I dilute it less the color in the ELISA is brownish instead of
>bright yellow (so way too dark).
>Is this normal??
>
>Suzanne
>
>
That sounds awfully high. 

 Are you certain that you have done all the usual
things like coating the plate with a non-reactive protein, adding  protein
to the buffers to fill nonspecific binding sites and so on?
That you are not using an excessive quantity of  second antibody, incubating 
too long, using fresh reagents  and so on.

OTOH, you seem to think you do get a reasonable curve.

I'm not an expert on these assays, but I would think that if you post
more specifics here, the antigen; immunization scheme; coating procedure;
elisa procedure  core info etc., someone will find an answer for you.







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