Add 200 l of TMB solution to each well with a multichannel pipette. Wells are very full. Add 50 l of stop solution with a multichannel pipette. Be sure all wells have turned yellow. If green color remains, continue mixing until green color turns to yellow.
Wells are very full.
Read in a plate reader at 450 nm. We recommend using a 4 parameter sigmoid minus curve fit. Some kit reagents contain small amounts of sulfuric acid, tetramethylbenzidine, acetone, methanol, or dimethyl sulfoxide, which as concentrated solutions may be toxic by inhalation, ingestion, or by contact with skin. Keep containers tightly closed and away from any sources of ignition. We recommend the procedures listed below for all kit reagents.