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Recently, many customers have reached out with questions about the washing process in ELISA kits. It's quite common for users to make mistakes during this step, which can significantly affect the accuracy of their results. At Shanghai Hengyuan, our experienced technical team has spent years working with ELISA kits and is here to share some essential tips to help you improve your experimental outcomes.
The washing step is crucial because it removes unbound antibodies that could cause background signals, thus improving the signal-to-noise ratio. Proper washing after each incubation ensures only specific binding events remain, leading to more reliable final results. If washing is not done correctly, you may end up with high background noise or inconsistent readings—both of which are undesirable.
When using an automatic washer, two key factors influence the effectiveness of the wash: the volume of wash solution used and the number of wash cycles. While increasing the number of washes generally reduces background, too many can also weaken the signal, making it harder to detect. A typical recommendation is to perform three washes after each antibody or antigen incubation. However, it’s always a good idea to check the manufacturer’s guidelines, as some ELISA plates may require fewer washes than others.
Another important point is the volume of washing liquid. For a standard 96-well plate, each well usually holds between 330–460 µL. Some automated washers allow you to program a higher volume, such as 1 mL, which can enhance the washing efficiency. The trick here is to use both the dispensing and aspiration functions simultaneously. This way, more liquid is added, but it doesn’t overflow into adjacent wells.
If you're still unsure about how to optimize your ELISA washing procedure, don't hesitate to reach out. Shanghai Hengyuan offers free testing and full technical support to help you get the best results from your ELISA experiments. For more information on ELISA technology, feel free to contact our sales team anytime. We’re here to help! Heng Jiaxing AO3401 Test - lowercase jpg Kaixin micro test Test probe P100-M3