lDilutesamples1:1distribing50ulofseruminto50ulofdilluent,Add50ulofdilutedsampletowells.lAdd50ulofdilutedbiotinylatedanti-porcineinsulintoallwells.lCoverwithaplatevoverandincubatefor1hourat37℃.lRemovethecoverandwashtheplateasfollows:⑴aspiratetheliquidfromeachwell,⑵dispensse0.3mlofwashingsolutionintoeachwell.⑶Aspirateagainthecontetofeachwellafter0.5minute.⑷Repeatsteps⑵and⑶threetimes.lDistribute80ulofstrept***idin-HRPsolutiontoallwells,includingblankwells.lCoverandincubate30minat37℃.lRemovethecoverandemptywells,Washmicrowellstripsaccordingtostep,Proceedimmediatelytothenextstep.lAdd50ulSubstrateAandSubstrateBtoeachwell。Incubatefor10minat37℃。lTheenzyme-substratereactionisstoppedbyquicklypipetting50ulofH2SO4.stopreagentintoeachwell,includingtheblankwells,tocompletelyanduniformlyinactivatetheenzyme.ResultsmustberedimmediatelyaftertheadditionofH2SO4.lReadabsorbanceofeachwellonaspectrophotometerusing450nmastheprimaryw***elengthandoptionally620nm(610nmto650nmisacceptable)asthereferencew***elength.说明书Email:info@索取;13801829628定货；HR-M0409)