Home Failure Case Library High CVs at Bottom of Standard Curve
ELISA (Competitive) moderate

High CVs at Bottom of Standard Curve

Symptom
Coefficient of variation (CV) among replicates is excessively high at the lowest standard concentrations (highest OD in competitive format), reducing precision at the sensitive end of the curve.
Common Causes
  1. 1 Higher OD values inherently demonstrate greater variance in competitive format
  2. 2 Pipetting errors more pronounced at small volume differences
  3. 3 Standard concentrations approaching detection limit of the assay
  4. 4 Samples falling outside optimal measurement range of curve
Solutions
  1. 1 Accept that higher ODs (lower concentrations) naturally show more variance in replicates
  2. 2 Increase the concentration range of the standard curve to shift measurement window
  3. 3 Dilute samples less so they fall in the middle of the standard curve
  4. 4 Use technical triplicates or quadruplicates at low concentrations to improve precision
Related Video (2)
Thermo Fisher Scientific ★ 75
How to Run an ELISA Assay – Invitrogen Kit Step-by-Step Tutorial
"Shows correct ELISA technique with emphasis on reproducibility, useful for troubleshooting CV issues."
Bilibili (China-Accessible Mirrors) ★ 70
How to Run an R&D Systems Quantikine ELISA
"Includes troubleshooting guidance within a complete Quantikine ELISA workflow, relevant to precision issues."
Source: abcam.com ↗
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