How can I determine the titre of my egg yolk antibody against bacteria..or can anyone plz tell me how to check antibody titre against bacteria.thank you?
Dear Chankit Giri,
You may start with simple agglutination assay, check the optimum concentration which can clump your bacterial strains.
If you have purified antibody you may fluorphore/HRP label it and try varying concentrations of it against known concentration of your bacterial antigen by ELISA.
Good luck,
Sabarinath.PS
Cover ELISA plates with log-phase bacteria, keep in 4 degree C, overnight. Then the next day, perform a simple indirect ELISA. As a primary antibody, you can use serum isolated from immunized animal. As a secondary antibody, use HRP-conjugated anti-mouse or anti-guinea pig or anti-rabbit antibody (depending on the animal from which you've isolated the serum). Then add a substrate and you'll get colored products. From which you can find out the titer value.
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