Operation steps of mouse interleukin 1β (IL-1β) ELISA kit:

Operation steps and precautions of mouse interleukin 1β ELISA kit

1. Sample addition: set up blank wells (the blank control wells do not add samples and enzyme reagents, the rest of the operation is the same), standard wells, sample wells to be tested. Add 50μl of the standard on the enzyme-coated plate accurately, add 40μl of sample diluent to the sample well, then add 10μl of the sample to be tested (the final dilution of the sample is 5 times) Add the sample and add the sample to the bottom of the well of the microplate, try not to touch the wall of the well, shake gently to mix.

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

3. Mixing solution: dilute 20 times concentrated washing solution with distilled water 20 times and reserve

4. Washing: Carefully peel off the sealing film, discard the liquid, spin dry, fill each well with the washing solution, let it stand for 30 seconds, then discard, repeat 5 times and pat dry.

5. Add enzyme: add 50μl of enzyme label reagent to each well, except blank well.

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

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

8. Color development: Add 50μl of developer A to each well, and then add 50μl of developer B, mix gently, and develop at 37 ° C in the dark for 15 minutes.

9. Stop: add 50μl of stop solution to each well to stop the reaction (at this time, the blue color turns to yellow).

10. Determination: Measure the absorbance (OD value) of each well with the blank air conditioner at zero and 450nm wavelength in sequence. The measurement should be carried out within 15 minutes after adding the stop solution.

Note for mouse interleukin 1β (IL-1β) ELISA kit:

1. The kit should be taken out of the refrigerated environment and equilibrated at room temperature for 15-30 minutes before use. If the enzyme-coated plate is unsealed after opening, the strip should be stored in a sealed bag.

2. Crystals may be precipitated in the concentrated washing liquid, which can be heated and dissolved in the water bath during dilution, and the results will not be affected during washing.

3. The sample adder should be used in each step of sample addition, and its accuracy should be regularly checked to avoid test errors. It is best to control the sampling time within 5 minutes. If there are many specimens, it is recommended to use a volley gun to add samples.

4. Please make the standard curve at the same time of each measurement, it is better to make the complex hole. If the content of the substance to be tested in the specimen is too high (the OD value of the sample is greater than the OD value of the first well of the standard well), please dilute it with a certain multiple of the sample diluent (n times) before measuring, and finally multiply by the total dilution Multiple (× n × 5).

5. The sealing film is limited to one-time use to avoid cross contamination.

6. Please keep the substrate away from light.

7. Strictly follow the instructions, and the test results must be determined based on the reading of the microplate reader.

8. All samples, washing liquid and various wastes should be treated as infectious agents.

9. The components of different batches of this reagent shall not be mixed.

10. If it is different from the English manual, the English manual shall prevail.

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