摘要：

2. Dispense 100 ml of standards and samples into the wells (we recommend that samples be tested in duplicate).3. Incubate on an orbital micro-plate shaker at 100-150 rpm at room temperature (18-25°C) for 45 minutes.4. Aspirate the contents of the microtiter wells and wash the wells 5 times with 1x wash solution using a plate washer (400 ml/well). The entire wash procedure should be performed as quickly as possible.5. Strike the wells sharply onto absorbent paper or paper towels to remove all residual droplets.6. Add 100 ml of enzyme conjugate reagent into each well.7. Incubate on an orbital micro-plate shaker at 100-150 rpm at room temperature (18-25°C) for 45 minutes.8. Wash as detailed in 4 and 5 above.9. Dispense 100 ml of TMB Reagent into each well.10. Gently mix on an orbital micro-plate shaker at 100-150 rpm at room temperature (18-25°C) for 20 minutes.11. Stop the reaction by adding 100 ml of Stop Solution to each well.12. Gently mix. It is important to make sure that all the blue color changes to yellow.13. Read the optical density at 450 nm with a microtiter plate reader within 15 minutes.

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