Hey,
we are currently working on a project in which we test for serum antibody responses against several different antigens, using a multiplex immunoassay. Our idea is to test for IgA, IgM and IgG simultaneously. In pilots, we compared the strength of IgA and IgG responses (in various dilutions) when run separately and when combined in one well. It seems like the secondary antibodies interact with each other, reducing the signal strength of both immunogolbulins.
Both secondary antibodies are goat anti-human and we are wondering if these interactions can be reduced if we use anti-human secondary antibodies produced in different species?
It would also be of interest if someone tried something similar before and has some experience or knowledge about secondary antibody interactions in general.
Thank you very much,
Marvin
Have you tried running your goat anti-human IgG against human IgA and vice versa. If there is any cross reactivity of the secondary antibodies then the reagents might work well alone, but could interfere with each other if used together. I would look into this and find anti-IgA/G/M reagents with very high specificity.
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