In vitro diagnostic (IVD) products

 

 

 

Step 1

Adding Negative Control, Test Sample and Blank:

Add 50μl of Negative Control, 100μl of test sample and 100μl of 1 x Assay Buffer as Blank into their respective wells. Triplicate test is recommended for Negative Control and duplicate test is recommended for Blank and test samples.

Note: Use a separate disposal pipette tip for each test sample, Negative

Control and Blank to avoid cross-contamination. Mix by tapping the plate gently.

Step 2 Incubation:

Cover the plate and incubate at room temperature for 1 hour.

 

 

Step 3

Washing:

Discard the content and tap the plate on a clean paper towel to remove residual solution in each well. Add 300 µl of 1×Wash buffer to each well and incubate for 1 minute. Discard the 1×Wash buffer and tap the plate on a clean paper towel to remove residual wash buffer. Repeat the wash step for a total  3 washes.

Step 4 Adding HRP-conjugated Detection Solution:

Add 100 µl of 1×Detection Solution to each well.

Step 5 Incubation:

Cover the plate and incubate at room temperature for 1 hour.

Step 6 Washing:

Wash each well 4 times as described in step 3.

 

Step 7

Colouring:

Add 100 µl of Substrate solution to each well,  incubate  at  room  temperature for 15 minutes. Protect from light.

 

Step 8

Stopping Reaction:

Add 100 µl of Stop solution to each well, gently tap the plate frame for a few seconds to ensure thorough mixing.

 

Step 9

Measurement:

Measure absorbance of each well at 450 nm immediately.

Note: read the absorbance within 10 minutes after stopping the reaction.