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QUICK ANSWER If your ELISA returns sample concentrations that are orders of magnitude too high, the assay itself is usually fine — the error is almost always in data analysis. The most common culprit is entering the wrong standard concentrations when building the standard curve. In a recent porcine MAP (PigMAP) sandwich ELISA case, a researcher assigned a 5000–78 ng/mL range to standards that should have been 10–0.156 ng/mL. The optical density (OD) readings were valid; only the concentrations tied to them were wrong. Re-assigning the correct values to the same ODs brought results back into a sensible range. On this page The problem Snapshot of t …

Quick answer On the GDF-6 (Growth Differentiation Factor 6) ELISA Kit, a researcher saw an elevated blank (OD ≈ 0.41) with the two lowest standards reading at or below it, and no signal for a recombinant GDF-6 sample. The cause was a background/handling issue — most likely an over-concentrated HRP conjugate, incomplete washing, or stale wash buffer — not a fault in the antigen or kit. Fresh reagents, thorough washing, and a rebuilt low-end curve resolve it. On this page Click any section below to jump straight to it ↓ The problem › Snapshot of the data › Why it happens › Our a …