Procedure for porcine major acute phase protein (MAP) test kit
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1. Adding samples: Set blank holes separately (the blank control wells are not added with samples and enzyme-labeled reagents, and the other steps are the same), standard wells, and sample wells to be tested. Accurately load 50 μl of the standard on the enzyme-labeled plate, add 40 μl of the sample dilution to the well 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 microplate, try not to touch the wall of the well, and shake gently to mix.
2. Incubation: After sealing with a sealing film, incubate at 37 ° C for 30 minutes.
3, dosing: 30 times concentrated washing solution diluted with distilled water 30 times, spare
4, washing: carefully remove the sealing film, discard the liquid, dries, fill each hole with the washing liquid, let stand for 30 seconds and then discard, so
Repeat 5 times and pat dry.
5. Add enzyme: Add 50 μl of enzyme labeling reagent to each well, except for blank wells.
6. Color development: first add color developer A50μl to each well, then add color developer B50μl, gently shake and mix, avoid light and develop color at 37°C
10 minutes.
7. Termination: 50 μl of stop solution was added to each well to terminate the reaction (in this case, the blue color turned yellow).
8. Measurement: The absorbance (OD value) of each well was measured in sequence with a blank air conditioner of zero and a wavelength of 450 nm. The measurement should be carried out within 15 minutes after the addition of the stop solution.