Product Details

Product Description

1. Bring reagents to room temperature (15-30°C) for at least 30 minutes before use.

2. Allow for eight Blank/Control determinations (two Reagent Blank, two Negative Control, two Low Positive Control and two Positive Control) per run.

3. A full or partial plate may be run. Store the residual strips from a partial plate run in the original pouch and seal the entire opening with adhesive tape. Do not use broken strips.

4. Determine the volume of 1X Wash Buffer needed for the run. 24 mL total of 1X Wash Buffer is needed for each full strip of controls and/or samples run. Volume to be prepared is (24 mL x # strips) plus sufficient excess to ensure adequate quantity for use with manual or automated wash. To prepare a 1X Wash Buffer, mix 1 part of the 20X Wash Buffer with 19 parts of purified water thoroughly in an appropriately sized container to achieve homogeneity. The 1X Wash Buffer may be stored at 2-8°C for up to 30 days.

5. For control wells, add 50 μL control solution directly to individual wells (except for the two Reagent Blank wells).

6. For sample wells, add 50 μL sample serum to each well.

7. Mix well by gentle shaking, cover the microplate wells with an adhesive sealer, and incubate for 30 minutes at 37±2°C.

8. Microplate Washing: discard the liquid in the wells then fill each microwell with 300 μL of working Wash Buffer (1X). Leave the wash buffer in each well for 5 seconds and then remove the wash solution from all the wells. Repeat 4 times for a total of 5 washes. Firmly tap the inverted microplate on absorbent paper to thoroughly remove all residual liquid from the wells. The preceding manual wash may be performed with an automated plate washer.

9. Add 100 μL of the Enzyme Conjugate to each well at the same rate and in the same order as the samples. Cover the microplate with an adhesive sealer and incubate for 30 minutes at 37±2°C.

10. Microplate Washing: wash the microwells by following the procedure as described in step 8.

11. Add 50 μL each (100 μL total) of Chromogenic Reagent A and Chromogenic Reagent B, to each well at the same rate and in the same order as the addition of the patient specimens. Mix well by gentle shaking, cover the microplate with an adhesive sealer, and incubate for 10 minutes at 37±2°C protected from direct sunlight.

12. Add 50 μL of Stop Solution to each well at the same rate and in the same order as the Chromogenic Reagents. Gently tap the microplate several times to ensure that the reagents are thoroughly mixed.

13. Set the microwell reader to read at a wavelength of 450nm and measure the optical density (OD) of each well against the mean of two Reagent Blank wells. Read the microplate within a maximum of 10 minutes of the addition of the Stop Solution.