Three Maxisorb 96-well plates from the same sleeve produced identical patterns of high absorbancies in wells that should not have had such signals. All 3 plates were double sandwich ELISAs. One plate was for IFNalpha (using SuperBlock as blocker), one for IFNbeta (using casein as blocker), and one for IFNlambda (using casein as blocker). We are using TMB as the coloration substrate for HRP. We've seen this before, but not with as many wells as this time.
Thanks for the response, Josh.
We adsorb primary monoclonal antibody to wells, wash, add block, wash, adsorb sample, wash, adsorb secondary polyclonal antibody conjugated to biotin, wash, adsorb streptavidin-HRP, wash, add TMB+H2O2, stop, read absorbance @ 465nm.
We have not reached out to the plate manufacture yet, but it is a good idea. We've seen this before in isolated wells, but never to the extent that we experienced during the last assay run.
Hi. Are you using automated ELISA equipment? If this is the case it is possible that the channels corresponding to the specific wells are not properly working.
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