ELISA Troubleshooting Well-to-Well Variation

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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.

ProblemCauseSolution
Plate handling problemsPlates stacked during incubationDo not stack plates to ensure even temperature distribution
Plates warmed unevenly before useAllow plate temperature to stabilize before use to avoid edge effects
Bubbles in wellsCheck for bubbles before incubations
Uneven washingWash all wells uniformly
Check plate washer for obstructed ports
Reagent problemsIncomplete reagent mixingMake sure all reagents are homogenous before use
Inconsistent storageMake sure all samples and reagents are stored under uniform conditions

Keywords:- elisa, troubleshooting, well-to-well, variation, high variation, intra-assay variation