AEC Peroxidase Substrate Kit From Enzo for Focus Forming Assay

This kit was used for focus focus-forming assay to determine the viral titer of the Dengue virus. For this assay, I followed the supplier's recommendation for reaction reagent preparation. For one 24-well plate, the following dilution was prepared: 312.5 uL of 8X reaction buffer + 2187.5 uL of distilled water, and only one drop of concentrate Aminoethylcarbazole was added to the mix. It is best to protect the mix from the light, so I prepared this solution in a 50 mL falcon tube covered in aluminum foil. I added 100 uL per well and incubated at room temperature for about 5 min until the precipitate was visible enough. The plate was kept covered in aluminum foil. Following precipitate visualization, the reaction was interrupted by washing wells with distilled water.