Chicken HSP60 Antibody (IgM) Kit Operation Method

Chicken HSP60 Antibody (IgM) Kit Operation Method

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Steps

1. Dilution of Standard: This kit provides one original standard. Users can dilute in small test tubes according to the following chart.

2. Adding samples: Set blank wells (the blank control wells do not add samples and enzyme-labeled reagents, the rest of the steps are the same), standard wells, and sample wells. Add 50 μl of the standard accurately to the enzyme-coated plate. Add 40 μl of the sample dilution to the wells of the sample to be tested, and then add 10 μl of the sample to be tested (the final dilution of the sample is 5 times). Add the sample to the bottom of the well of the microtiter plate, try not to touch the wall of the well, and shake gently to mix.

3. Incubation: Seal the plate with a sealing plate and incubate at 37 ° C for 30 minutes.

4. Dosing: Dilute 30 times concentrated washing solution with distilled water 30 times and set aside.

5. Washing: Carefully peel off the sealing plate film, discard the liquid, and dry it. Fill each well with washing liquid, leave it for 30 seconds, and discard it. Repeat this 5 times and pat dry.

6. Add enzyme: Add 50μl of enzyme-labeled reagent to each well, except for blank wells.

7. Incubation: The operation is the same as 3.

8. Washing: The operation is the same as 5.

9. Color development: Add the color developer A50μl to each well, and then add the color developer B50μl, shake gently to mix, and develop the color at 37 ° C in the dark for 15 minutes.

10. Termination: Add 50 μl of stop solution to each well to stop the reaction (the blue color turns to yellow at this time).

11. Measurement: Measure the absorbance (OD value) of each well sequentially with a blank air conditioner at a wavelength of 450nm. The measurement should be performed within 15 minutes after adding the stop solution.