Dear Kevin, I think that if we do not wash the biotinylated conjugate before adding the streptavidin (SA), we probably need more SA since the excess of biotin-conjugate can remove a lot of unbound SA and your system will become more expensive. Moreover, in cases where you have a little binding of your biotin-conjugate to the target, the competition with the unbound ones could generate false negative results. Best regards, Olindo
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