Shanghai Jinma narrator human interleukin-1 ELISA kit operation steps - Database & Sql Blog Articles

Shanghai Hengyuan Biotechnology Co., Ltd. newly launched the human interleukin-1 (IL-1) elisa kit in October 2015. The students of Shanghai Tongji University did not use the product for the first time. Familiar with, consult our technical consultants, the company's technician Ouyang professor answered this question for the first time, welcome the industry to refer to. The specific procedure of human interleukin 1 (IL-1) elisa kit is 11 points, as shown below:
1. Dilution of standard: This kit provides one original standard, which can be diluted in a small tube according to the following chart.
Add 150 μl of standard dilution to 240 ng/L No. 5 standard 150 μl of the original standard
120ng/L No. 4 Standard 150μl of No. 5 Standard Add 150μl Standard Diluent
60ng/L No. 3 Standard 150μl Standard No. 4 Add 150μl Standard Diluent
Add 30 μl standard dilution to 30 ng/L No. 2 standard 150 μl No. 3 standard
Add 150 μl of standard dilution to 15 ng/L No. 1 standard 150 μl No. 2 standard
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), the standard holes, and the sample holes 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.
3. Incubation: The plate was sealed with a sealing film and incubated at 37 ° C for 30 minutes.
4. Dosing: Dilute 30 times concentrated washing solution with distilled water 30 times.
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: The operation is the same as 3.
8. Wash: Operate the same as 5.
9. Color development: Add 50 μl of the developer to each well, then add 50 μl of the developer, gently shake and mix, and develop at 37 ° C for 15 minutes.
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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