1.Prepare all reagents, samples and standards;
2.Add 100uL standard or sample to each well.Incubate 2 hours at 37℃;
3.Aspirate and add 100uL prepared Detection Reagent A.Incubate 1 hour at 37℃;
4.Aspirate and wash 3 times;
5.Add 100uL prepared Detection Reagent B.Incubate 30 minutes at 37℃;
6.Aspirate and wash 5 times;
7.Add 90uL Substrate Solution.Incubate 10-20 minutes at 37℃;
8.Add 50uL Stop Solution.Read at 450nm immediately.
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