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	Can a conjugate antibody bind to a well that is not coated with a capture antibody? An empty well not coated with antibody, but did get blocked, went through the steps of conjugate and substrate and was bright yellow, even though the well was never coated and never had a sample in it. Can anyone help explain?
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	Hi there!
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	Strong signal in negative wells is a sign of unspecific binding. The blocking may not be complete so that the conjugate binds to the well or there is crossreactivity between the conjugate and some substance in the blocking solution. Which antibodies/system was used?
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Colour development in a well with no coating antibody?