Is it possible to use HRP-conjugated secondary Ab (for detection Ab in sandwich ELISA) without using the biotin-avidin?

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Viktor Y Butnev

Yes, it's possible, but only if all your three antibodies (capture Ab, 1st antibody, and 2nd antibody-HRP) will belong to different species and will not show any cross-reactivity.

Gertrudis Rojas

Of course you dont need the biotin/streptavidin system. Actually the coating antibody and the conjugated antibody can belong to the same species. You need to take care that there are not undesired crossreactivities, as Viktor suggested.