Pre-coated, ready to use 96-well strip plate Standard (freeze dried) Standard Diluent Detection Reagent A Detection Reagent B Assay Diluent A Assay Diluent B TMB Substhumane Stop Solution Wash Buffer(30 x concenthumane) Plate sealer for 96 wells Instruction manual
Material not included
1. Microplate reader with 450 ± 10nm filter. 2. Precision single or multi-channel pipettes and disposable tips. 3. Eppendorf Tubes for diluting samples. 4. Deionized or distilled water. 5. Absorbent paper for blotting the microtiter plate. 6. Container for Wash Solution.
Optimal working dilution should be determined by the investigator.
Assay Time
1 - 4.5 h
Plate
Pre-coated
Protocol
1. Prepare all reagents, samples and standards 2. Add 100µL standard or sample to each well. Incubate 2 hours at 37℃ 3. Aspirate and add 100µL prepared Detection Reagent A. Incubate 1 hour at 37℃ 4. Aspirate and wash 3 times 5. Add 100µL prepared Detection Reagent B. Incubate 1 hour at 37℃ 6. Aspirate and wash 5 times 7. Add 90µL Substrate Solution. Incubate 15-25 minutes at 37℃ 8. Add 50µL Stop Solution. Read at 450nm immediately.