If I immunize mice with a recombinant P. falciparum antigen and use ELISA to check the presence of antigen-specific antibodies, what will be my positive control antigen? I was thinking of using E.coli cell lysate.
Positive controls are highly dependent on the purpose of your work. If you are not sure, which positive control should be chosen, the best option is to use as many controls as you can. BTW: In your case, the most obvious test is the coating of purified antigen on a microtitration plate.
Hi Michael,
To check for the presence of the antigen-specific antibody in the mouse serum, after immunization, I will have to coat the plate with my test antigen and I am wondering what other antigen I could use as a positive control for this test.
Do you have the same antigen in non-recombinant form, e.g isolated from Plasmodium falciparum? You still need to define, what you want to do with your "antigen-specific" antibody. Is your target native or denatured, a living parasite or fixed? Otherwise, your antibody might not work under your anticipated assay conditions.
If you dont have any other source of your antigen, no positive control can be included. But a negative control antigen (one that should not be recognized by your sera unless there is non-specific background) should be included. Including the E.coli lysate (w/o teh recombinant antigen) is not a positive control because you tried to immunize with the purified antigen, I guess, so if there are contaminants proteins from E.coli that would be by accident .
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