Two Most Common Pitfalls To Avoid While Doing Elisa

ELISA is an easy and simple method for the quantitative or qualitative detection of peptides, proteins, antibodies and hormones in samples and is thus one of the most frequently used immunoassays. 

Despite the many advantages of doing ELISA, there are a number of mistakes that can quickly worsen your ELISA experiment. To know more about multiplex elisa visit

Read more about the 2 most common pitfalls you should avoid when doing ELISA to prevent this situation!

  1. “I think I still have some TMB stains from the previous kit. Maybe I’ll use that. “

Hold on! Demand for resource savings is a good thing, but not in this case. We must avoid using reagents from different lots at the same time. Each reagent in the kit is optimized for the lotto ensure the best performance. Using reagents from different lots could affect the ability to detect ELISA and lead to conflicting experimental results. In addition, previous reagents may have deteriorated depending on the manufacturing date and storage conditions.

Remember: Do not mix reagents from different lots.

  1. “Hmm … maybe there isn’t enough space in the incubator, so I’ll arrange this plate instead.”

Placing plates during incubation can have the final effect. Edge effects occur when external wells are exposed to different conditions (eg different temperatures) than neighboring internal wells, resulting in unexpected and conflicting values. The plates incubated unevenly resulted in higher CV values ​​for OD measurements.

To control for wells affected by edge effects, we recommend doubling or even multiplying the well analysis for standard and sample so that comparisons can be made if large differences occur.

Remember: For more consistent results, do not stack plates during incubation.