ELISA Basic Operation Steps Demonstration

critical

No Signal or Weak Signal in ELISA

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.

💡 6 · ✓ 6

critical

Poor or Non-Linear Standard Curve

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.

💡 5 · ✓ 5

severe

Inadequate Washing Between Assay Steps

High background with retained signal in negative controls. Edge wells may show higher background than center wells. Residual reagents visible in wells.

💡 4 · ✓ 5