Things to Be Aware of When Using ELISA Kits
When performing ELISA experiments, it is crucial to follow the manufacturer's instructions carefully. All results should be determined based on the readings obtained from a microplate reader to ensure accuracy and reliability.
If the enzyme-labeled reagent is not fully used after opening, it should be immediately placed back into a sealed bag with a desiccant to maintain its stability and effectiveness.
To minimize experimental errors, it is recommended to run all standards, samples, and blank controls in duplicate and take an average of the results. This helps improve the precision of your measurements.
Keep in mind that if your sample was diluted five times during the procedure, you must multiply the final result by 5 to obtain the actual concentration of the original sample.
The quantitative range of this kit is between 0.78 ng/L and 50 ng/L. If the measured value falls outside this range, the result can be estimated by extrapolating the standard curve, but this should not be considered as a precise measurement. To obtain accurate results, dilute the sample using a specific dilution solution and repeat the test within the valid range (0.78–50 ng/L). The final concentration is then calculated by multiplying the measured value by the total dilution factor.
If the color development is too light, you may extend the incubation time with the substrate slightly. However, be cautious not to over-incubate, as this could lead to inaccurate readings.
To prevent cross-contamination, use separate pipette tips for each standard, sample, and blank control. Avoid touching the microwell plate with shared reagents such as the enzyme standard working solution, sample diluent, or substrate. Also, do not reuse the sealing film, as this can introduce contamination.
Make sure to use the ELISA kit before its expiration date. Do not mix reagents from different batches, as this may affect the performance and consistency of the assay.
Substrate B is sensitive to light, so it should be stored and handled away from direct sunlight. Prolonged exposure to light can degrade the reagent and affect the results.
By following these guidelines, you can ensure more accurate and reproducible results when using ELISA kits. Always pay attention to details, maintain good lab practices, and stay consistent with your procedures.
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