Troubleshooting ELISA and EIA-based kits
Each ELISA or EIA kit includes a QC (quality control) certificate for quality control purposes. Adherence to the respective kit protocol should provide results that are similar to the QC data for each specific kit lot. In practice, local factors such as differences in ambient temperature, variations in pipetting equipment, variable plate washing characteristics, and inconsistent plate mixing and/or rotation may produce erratic or erroneous results. Below you will find some technical hints offered to help troubleshoot and optimize your EIA based assay kits.
I have poor precision between duplicates (high CV values).
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My standard curve is poor; it does not match the QC sheet.
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All my microplate wells turned bright blue!
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I have high background OD readings.
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I obtained unexpected results.
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There was a database error.