Real lab failures, root causes, and fixes — curated and bilingually annotated by our team.
Sample quantification values are outside expected biological range, controls fail to meet acceptance criteria, or results are not reproducible between runs despite similar experimental conditions.
The ELISA plate reader detects no colorimetric signal or signal intensity is significantly below expected values after substrate development. Wells appear clear or only faintly colored even after standard incubation times of 10-30 minutes.
Standard curve shows irregular shape, poor linearity, or inconsistent serial dilution response. R-squared value below 0.95 or curve does not follow expected sigmoidal or linear pattern across dynamic range.
Color intensity varies significantly between wells that should be identical. Pattern may show edge effects, gradients across the plate, or random spotty appearance indicating incomplete or inconsistent reagent contact.
Coefficient of variation (CV) between duplicate or triplicate wells exceeds 10-15%. Individual replicates show inconsistent absorbance values that cannot be attributed to biological variation.
All wells including negative controls show elevated absorbance readings. Background signal obscures specific signal, reducing signal-to-noise ratio and making data interpretation difficult.
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