| | Re: Problem in ELISA test
Post the protocol.
Coating antibody to the plate is simple enough, but quality of the buffer and the accurate pH is important, so fresh buffer is key.
Similarly the buffer for diluting the standard is also important as well as the mixing, I suggest gentle pipette mixing at first, to see how the standard protein performs.
Lastly the blocking step is critical, it blocks any sticky sites on the plate surface (plastic binds protein naturally, so empty spots need to be coated with inactive protein).
Provide more details if you want specific help.