Human creatine kinase brain isoenzyme (CK-BB) elisa technical specification Guangrui Bio- domestic high and excellent Elisa kit supplier steps: 1. Standard dilution and loading: on the enzyme label coated plate Set the standard hole 10 holes, add 100μl of the standard in the first and second holes, then add 50μl of the standard dilution in the first and second holes, mix; then from the first hole, the second hole Add 100 μl to each of the third and fourth wells, add 50 μl of the standard dilution to the third and fourth wells, and mix; then discard 50 μl in the third and fourth wells, respectively. Add 50 μl to the fifth and sixth wells respectively, then add 50 μl of the standard dilution solution to the fifth and sixth wells, and mix; after mixing, add 50 μl from each of the fifth and sixth wells. In the seventh and eighth holes, add 50 μl of the standard dilution solution to the seventh and eighth wells, mix 50 μl from the seventh and eighth holes, and add them to the ninth and tenth holes. 50 μl of the standard dilution solution was added to the ninth and tenth holes, and 50 μl of each of the ninth and tenth holes was discarded after being mixed. (The amount of each well was 50 μl after dilution, and the concentrations were 360 ​​μg/L, 240 μg/L, 120 μg/L, 60 μg/L, 30 μg/L, respectively). 2. Adding samples: Set blank holes separately (the blank control wells are not added with the sample and the enzyme standard reagent, the other steps are the same), and the sample holes to be tested. Add 40 μl of the sample dilution to the sample well to be tested on the plate, 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. 3. Incubation: After sealing with a sealing film, incubate at 37 ° C for 30 minutes. 4. Dosing: Dilute 30 (20 times of 48T) concentrated washing solution with distilled water 30 (20 times of 48T) and set aside. 5. Washing: Carefully remove the sealing film, discard the liquid, dry it, fill each well with the washing solution, let stand for 30 seconds, then discard it, repeat 5 times, and pat dry. 6. Add enzyme: Add 50 μl of enzyme labeling reagent to each well, except for blank wells. 7. Incubation: operation is the same as 3. 8. Washing: operation is the same as 5. 9. Color development: add 50 μl of color developer A, add 50 μl of color developer B, gently shake and mix, and avoid light at 37 °C. 15 min.10. Termination: 50 μl of stop solution was added to each well to stop the reaction (the blue color turned yellow). 11. Measurement: The absorbance (OD value) of each well was measured sequentially 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.

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