ELISA is a quite popular test. You need certain things for ELISA sample preparation such as serum saliva, milk etc. However, many times, ELISA test does not produce a good result if it is conducted in a wrong way. Therefore, you must be careful while conducting a test. Here are 3 ways of conducting the best ELISA test.
The key to run a successful assay is to have a low coefficient of variability between your duplicate samples. To keep %CV low between duplicates, you can use the pipet tips for every addition to the assay. Using new tips helps to prevent cross-contamination between wells which keep the %CV and background low and thus increase the reliability of the data. In ELISA sample preparation, this will really help to boost consistency and reliability.
To increase the accuracy, keep duplicated for all assay points such as standards, background and sample wells. If you run duplicates for all points, it will add to your good lab practice. Through this, you can immediately spot outliers that can affect your average. For researchers, more accurate data is important. This will give them confidence in their collected data and they can align their data with other published findings.
Produce Reliable Standard Curves:
In ELISA standard curve troubleshooting, to dilute the curve, you should use indicated range in the assay manual. Fresh standard curves should be made for every plate. There is a time limit for most assays for which fresh standard curve should be used. It is important to follow all precautions and recommendations for preparing a standard curve. For an appropriate troubleshooting, it is vital to include all suggested backgrounds wells in an assay.
To conduct ELISA Test, remember temperature, time, and shaking can affect the status of the assay if not followed properly.