(7)Add50 lperwellofthep-nitrophenylphosphatesolutionandincubateatroomtemperaturefor10-30min.Stopthereactionsbyadding50 lof0.1MEDTApH8.0.Forscreeningmonoclonalsitisbesttoletthemgountilallofthepositivesareclearlyvisible,howeverifyouwanttobequantitativeitisimportantnottoletthemgetintenselyyellowortheywillbeoffscale--theplatereaderismostaccuratewhenthewellshavejustbeguntoclearlyturnyellow.
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