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Is the sensitivity o...

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Is the sensitivity of ELISA kit low?


Recently, a customer reported that all the plate holes, including the positive control and quality control, had lighter colors after completing the experiment. What is the reason for this? After our technical organization, we provide the following analysis and invite you to take a look!


Possible causes include:
1. The reagent kit must be used within its expiration date, as products that have exceeded their expiration date may produce weak signals;
2. The reagent kit was not stored according to regulations and was affected by high temperatures;
3. Reagents and samples are not equilibrated to room temperature before use;
4. The volume and time of adding reagents are incorrect, the pipette measurement is inaccurate, and there is too much water or uncleanliness in the suction nozzle;
5. During the washing and sample addition process, the enzyme label becomes contaminated and deactivated, resulting in the loss of its ability to catalyze the color development of the color reagent;
6. The incubation time and temperature did not meet the requirements. When placing the reaction plate in the incubator, pay attention to the temperature and adjust it in a timely manner;
7. Wash the board too many times, or the dilution ratio of the concentrated washing solution does not meet the requirements; The washing impact is too strong. The soaking time is too long;
8. Insufficient amount or reversed order of color developer, or added after mixing;
9. Insufficient substrate action time;
10. There is a problem with the quality of distilled water.

terms of settlement:
1. Before the experiment, check the composition and batch number of the reagents to confirm that they have not expired.
2. Do not keep the reagent kit at room temperature for a long time and store it according to the usage regulations.
3. Ensure that the volume of reagents used is correct and the addition time is appropriate.
4. Calibrate the pipette and ensure that it fits tightly with the suction nozzle. The transfer should not be too fast, and the discharge should be complete.
5. Confirm that the container containing the enzyme label does not contain enzyme inhibitors such as sodium azide, confirm that the container used to prepare the washing solution has been cleaned, and confirm that the purified water used to prepare the washing solution meets the requirements and is not contaminated.
6. The incubation temperature should be controlled at 37-38 ℃, and the incubation time should be strictly operated according to the instructions. During the insulation period, it is not advisable to open more doors to avoid affecting the insulation.
7. Dilute and concentrate the washing solution and wash the plate strictly according to the instructions, reduce the washing impact force, and keep the washing solution for the required time and number of washes according to the instructions.
8. The color developer dropper bottle should be vertically downward with even holding force, and the dripping speed should not be too fast. First add color developer A, then add color developer B. Do not mix liquid A and B before adding
9. Accurate timing.
10. Determine the effect of distilled water preparation reagents on enzyme immunoassay.

Shanghai Hengyuan Biotechnology specializes in providing technical services in the field of life sciences, offering comprehensive experimental technology services, greatly saving the time and energy of repeated labor for scientific researchers. We have excellent technical consultation and perfect after-sales service. Purchasing ELISA reagent kits comes with free testing services, online one-on-one technical guidance, and can solve your worries.

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