ELISA kits are widely used in biomedical research and diagnostic testing, but the quality of reagents plays a crucial role in ensuring accurate results. Different manufacturers may produce reagents with varying levels of sensitivity and specificity. Using low-quality or expired reagents can lead to false positives or false negatives, which can significantly impact the reliability of your experiments. Choosing high-quality ELISA reagents is essential for achieving consistent and reliable outcomes. In this article, we’ll walk you through common issues that arise during ELISA experiments and provide practical solutions to help you avoid them.
| Steps | Possible Causes | Solutions |
| Reagent Selection | Selecting low-quality reagents or not following instructions properly. | Choose high-quality reagents, follow manufacturer guidelines, and allow reagents to reach room temperature before use (about 30–60 minutes). |
| Loading | Improper sample handling, delayed incubation, or enzyme splashing. | Ensure proper sample preparation, load samples promptly, and dry the plate surface after adding the enzyme reagent. Use automated systems if possible and work in batches when needed. |
| Incubation | Incomplete sealing or extended incubation time leading to non-specific binding. | Seal plates properly and strictly follow recommended incubation times. |
| Washing | Inadequate washing due to manual errors or machine inefficiencies. | Ensure full coverage of wash buffer, check for clogs, and dry the plate gently after washing. Use multiple washing machines if necessary. |
| Color Development | Expired developer or improper addition leading to contamination. | Prepare the developer fresh before use, avoid contact with metal, and ensure no overflow during application. |
| Stop Solution | Bubbles introduced during stop solution addition may cause false positives. | Add the stop solution carefully to avoid air bubbles. |
| Reading | Contaminants on the plate bottom can affect accuracy. | Clean the plate thoroughly before reading. |
| Overall Process | Exposure to contaminants or lack of standardization. | Keep the microplate away from contaminants and consider automation to improve consistency and quality. |
During actual operation, it’s not only important to choose high-quality reagents but also to strictly follow all procedural steps. Implementing internal quality control and inter-laboratory comparisons can further enhance the accuracy and reliability of your results. Maintaining a professional and meticulous approach to each specimen is key to ensuring high-quality testing. With the increasing adoption of automated microplate readers in many laboratories across China, the standardization and efficiency of ELISA testing have significantly improved. If you're looking for reliable and cost-effective ELISA kits, we’re here to help. Contact us via phone or QQ for more information. Our ELISA kits come in 96T and 48T formats, suitable for various experimental needs.
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