ELISA - coating antibody (Feb/02/2008 )
i a working on the antibody based ELISA
so plese say query is how the antibody raised from the pure protein .
and what type of protein require for that
It depends on your protein and your antibodies.
The first thing you should to do is an ELISA with different dilutions of antigen, and a constant dilution of the primary and secondary antibody (ELISA to determine the optimal dilution of your antigen).
A second ELISA, with dilutions of the primary antibody and antigen constants and different dilutions of secondary (to determine the optimal dilution of your secondary antibody).
Finally, an ELISA with constant dilutions of antigen and secondary antibody and different dilutions of your primary antibody to know the title of your antibody.
you may want to check these wikipedia pages :
antibodies
and
polyclonal antibodies
and
monoclonal antibodies
they give brief descriptions of antibody production.
so plese say query is how the antibody raised from the pure protein .
and what type of protein require for that
as for my knowldge most of the ELISA s based on antibodies.if you want to develop an antibody against you proteins , first you should know about ,whether your protein is strong immunogen or not in selected animal.For monoclonal and polyclonal antibody production you need to have more than 90%pure,otherwise you will endup with coross reacting antibodies.