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**Technical Notes for the Swine Encephalitis Antibody (JE-Ab) ELISA Method**
1. Before starting the test, remove the ELISA kit from the refrigerator and allow it to reach room temperature for 15–30 minutes. If the enzyme-labeled strip is not immediately used after opening, store it in a sealed bag to prevent moisture exposure and maintain stability.
2. The concentrated washing buffer may crystallize during storage. When preparing the working solution, gently heat it in a water bath to dissolve any crystals. This process does not affect the performance or accuracy of the assay.
3. Use a dedicated pipette for each step of the procedure to ensure precision. Regularly check and calibrate the pipette to avoid measurement errors. It is recommended to complete the sample addition within 5 minutes. For large sample volumes, using an automated pipetting device is highly advised to improve efficiency and consistency.
4. Always prepare a standard curve for each run, and it is strongly recommended to perform duplicate measurements for each standard. If the OD value of the sample exceeds that of the first standard well, dilute the sample with a suitable dilution factor (e.g., ×n×5) before retesting to ensure accurate quantification.
5. The sealing film should be used only once per plate to prevent cross-contamination between different samples or standards.
6. Protect the substrate solution from direct light exposure, as it can degrade over time and affect the reaction outcome.
7. Follow the manufacturer’s instructions carefully. All results must be interpreted based on the microplate reader readings, not visual observations.
8. Treat all samples, washes, and waste materials as biohazardous substances. Proper disposal procedures should be followed according to local regulations.
9. Do not mix components from different batches of the reagent kit, as this may lead to inconsistent or unreliable results.
10. In case of discrepancies between the English and other language versions of the manual, the English version shall take precedence.
By following these guidelines, you can ensure the accuracy, reliability, and reproducibility of your JE-Ab ELISA results. Always maintain a clean and organized work area, and keep detailed records of your experimental conditions and data.
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