an ELISA procedure for Measuring murine serum Ab titer - does anybody know of such an ELISA protocol or kit? (Apr/30/2007 )

Hey guys, long time since I last visited here...

Anyway, my boss wants to set up a system of ELISAs for detecting serum Ab titers following immunization of mice with some of the proteins we work with.

I was trying to find a protocol for such an ELISA or a kit for it and came out with one. I would appriciate it if u could enrich my knowlege and sher it with me if u know of such a protocol.

Thanks.

Amir.

-taliasa-

QUOTE (taliasa @ May 1 2007, 01:45 AM)

Hey guys, long time since I last visited here...

Anyway, my boss wants to set up a system of ELISAs for detecting serum Ab titers following immunization of mice with some of the proteins we work with.

I was trying to find a protocol for such an ELISA or a kit for it and came out with one. I would appriciate it if u could enrich my knowlege and sher it with me if u know of such a protocol.

Thanks.

Amir.

Coat your plate with anti IgG (or IgM or whatever; say sheep anti-human), then detect with anti Ig (say rabbit anti-human). Use two different organisms for the two antibodies, so you get minimal crossreaction.
Alternately, you could coat with protein A or protein G, and use just 1 anti-Ig Ab (make sure it's an enzyme conjugate, or course).

-swanny-

Or coat your immunogen protein or peptide onto the plate and titrate your murine serum antibody on that and detect with an anti-mouse enzyme conjugate.

-WAstate-