Recombinant antigen-Elisa - (Nov/29/2006 )

Hi everybody;

I'm new in this world of Elisa, and I don't have any experience in it, so I have a lot of question....

I cloning a protein of virus HIV in E. COli and I want to know if this protein has inmunological activity, so I decided to design an Elisa, I will coat my recombinant protein and then I will prove several human serum of patients to check if my protein could differentiate between positive seum and negative. The QUESTION is.... which is the first step to do?

Thanks a lot for your collaboration

Ana.

Have you already done chessboard/checkerboard titration (CBT)? (ie. diluting antigen by columns and antibody by rows (or vice versa), used to optimize conditions of ELISA)