I am not seeing any signal after ELISPOT assay and wondering if I can re-probe.
It is possible depending on how the assay was carried out. If all the steps were carried out correctly but a wrong species conjugate was used, simply wash the plate, dry and use the correct conjugate. ((I tried this with ELISA; used the wrong species conjugate, washed and then used the correct species conjugate and got good signals)).
If all the steps of the protocol were carried out "correctly' and you got no signal, then there's not much you can do. In this case, I will suggest you mentally go through all what you did to see if you used the right reagents. If a correct reagent was used at a sub-standard concentration, repeat the experiment with a new plate.
If the failed experiment just ended and you realized you performed a step wrongly, you can try your luck by washing the plate and continue from the step where the error was made. After all, we all know that (I) these plates are expensive and (II) anything not specific for that assay is not suppose to bind.
Good luck !
I agree with Jerome. I have used the wrong secondary in an elispot, washed, reprobed and got great results
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