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For the teachers who are doing elisa experiments, is the success of elisa? There are a lot of tricks. These tips seem trivial, but they actually play a vital role in the experiment. Master these tips. Allows you to be more proficient in the elisa experiment, greatly increasing the chances of success in the elisa experiment.
The 17 related operation skills of the ELISA kit are as follows:
1. Remember to use a blotting paper to gently blot and dry on the surface of the microplate after adding the enzyme reagent.
2. Arrange the amount of detection reasonably so as to avoid long waiting time for washing the plate.
3. When sucking the liquid, it is necessary to remove the gun with a dose and a quantity close to the required amount to reduce the error.
4. Be sure to do a two-hole experiment so that the accuracy of the data can be guaranteed and the precision of the kit can be reflected.
5. The sample diluent is filled with a pipette and often corrected for accuracy.
6. In order to prevent the sample from evaporating, the reaction plate is placed in a closed box covered with a damp cloth, and the plate is covered with a cover or a film.
7. Unused plate or reagent, please store at 2-8 °C. Horseradish peroxidase-labeled anti-human IgG working solution should be used according to the required amount. Do not reuse the diluted horseradish peroxidase-labeled anti-human IgG working solution.
8. In the ELISA kit experiment, the incubator should be placed in time after the sample is added. When there are more specimens, it must be operated in batches. According to the instructions, the operation time is strictly controlled to prevent the incubation time from being artificially prolonged, resulting in non-specific binding surrounding the reaction well, which is difficult to clean thoroughly.
9. The remaining samples and waste should be sterilized by autoclaving at 121 °C for 30 minutes, or treated with a disinfectant such as 5.0 g/L sodium hypochlorite for 30 minutes.
10. When the human ELISA kit is washed, each well needs to be filled with liquid to prevent the free enzyme in the orifice from being washed.
11. One hole was left for each experiment as a blanking zero hole. No additional reagent was added to the well, except that the substrate solution and 2N H2SO4 were added. Use this hole to adjust the OD value to zero when measuring.
12. After washing the plate manually, add the washing solution each time. Should be allowed to stand for 15 ~ 30s, do not splash a washing solution in the enzyme label L into another enzyme labeled hole to prevent cross-contamination. After removing the washing solution, place the microplate on a towel or absorbent paper and pat dry.
13. When there is doubt about the results of the sample, other test methods need to be used for verification.
14. When there is no deionized water or double distilled water, you can use Wahaha pure water to prepare the solution. Do not use tap water.
15. When using the micro-sampler, replace the tip with the liquid in different bottles, even if the standard solution is taken.
16. When the liquid is sucked, the speed is not too fast, so as to avoid air bubbles, and the amount of the human ELISA kit is not accurate enough.
17. When sucking the liquid, use a micro-sampler with a close range and a required amount to reduce the error.
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