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