Hengyuan tells you how to reduce the error probability of ELISA kits - Database & Sql Blog Articles

How can the experimental error probability of an ELISA kit be reduced? In addition to being more careful during operation, there are several key points that should not be overlooked. Shanghai Hengyuan Bio has summarized nine important aspects for you: △ Aspect 1: The technician performing the test should have strong laboratory skills and hands-on experience. They should be familiar with operating lab equipment and possess the ability to analyze and resolve issues quickly when they arise. △ Aspect 2: Use a calibrated micropipette to minimize human errors. The accuracy of the pipette is crucial, especially in quantitative assays where even small deviations can affect results. △ Aspect 3: Always read the instructions carefully and follow them precisely. Do not mix reagents from different batches of ELISA kits. If improvements are needed, conduct trial and error tests after establishing a stable protocol. △ Aspect 4: Ensure thorough washing of the microplate. Incomplete washing may lead to a high background signal, resulting in false positives. Use fresh wash buffer and avoid reusing old solutions, which can increase background noise. △ Aspect 5: Control the reaction time strictly. Too long a reaction may inactivate the enzyme, while too short a time may prevent complete binding between the conjugate and the antigen-antibody complex, leading to false negatives. △ Aspect 6: Pay attention to the shelf life of the ELISA kit. Reagents past their expiration date should not be used, as they may no longer perform reliably. △ Aspect 7: Evaluate the performance of the reagent before use. Conduct control experiments with positive and negative samples to ensure the reagent is functioning properly. Only use it if it meets the required standards. △ Aspect 8: Monitor the color development time carefully. If it’s too short, the reaction may terminate prematurely, causing false negatives. Conversely, if it's too long, it may result in false positives due to excessive substrate accumulation. Always observe the color change promptly and stop the reaction at the right time. △ Aspect 9: Handle the sample accurately and efficiently. Inaccurate sample handling can affect the enzyme activity and ultimately impact the final results. The volume of the developer and stop solution added also influences the A value, so all steps must be performed with precision. If the sample is slow to process or left exposed for too long, especially in high temperatures, the reagent may degrade, leading to unreliable results.

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