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Troubleshooting guides

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ELISA Troubleshooting: High Variation

High well-to-well variation can be a symptom of a flawed experiment, or it can be a simple error that’s easy to fix. Errors in plate handling or loading, or mistakes in the usage of reagents are all common causes of well variance in ELISA. Use this troubleshooting guide to help solve some of the common sources of error that drive high well-to-well variability.

Problem Cause Solution
Plate handling problems Plates stacked during incubation Do not stack plates to ensure even temperature distribution
Plates warmed unevenly before use Allow plate temperature to stabilize before use to avoid edge effects
Bubbles in wells Check for bubbles before incubations
Uneven washing Wash all wells uniformly
Check plate washer for obstructed ports
Reagent problems Incomplete reagent mixing Make sure all reagents are homogenous before use
Inconsistent storage Make sure all samples and reagents are stored under uniform conditions

Keywords:ELISA, Troubleshooting, Variation, Well Variation

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