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ELISA - antibody mixture - (Sep/29/2015 )



I have incubated my 96-well elisa plate overnight. I just realise i made a wrong mixture. I put 1ul of antibody along with 20ml of elisa buffer. Supposedly 1ul of antibody for 25ml of elisa buffer. Will there be any effect? I will block my plate tomorrow



Any effect is always antibody/analyte dependent but it will likely be fine.  You are just 25% more concentrated than the instructions intended.  This may have no effect or it could increase the sensitivity of the assay.  Depending on the antibody/assay, it also could increase background signal but it probably won't.