Briefly steps:
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1. In each hole adding standard or sample 100 mu L, 37 ℃ for 90 minutes incubation
2. Add in 100 mu L biotin antibody working liquid, 37 ℃ incubation 60 minutes
3. Wash three times
4. Add in 100 mu L enzyme combined with the working liquid, 37 ℃ incubation for 30 minutes
5. Washing five times
6. Join the 90 mu L substrate solution, 37 ℃ incubation for about 15 minutes
7. Add 50 mu L terminated liquid, at 450 nm wavelength measuring OD value immediately
8. The results calculated
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