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Troubleshooting guides

Troubleshooting guides

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ELISA Troubleshooting: Low Sensitivity

When using small amounts of precious protein sample, the sensitivity of your ELISA assay can be its most important feature. Boster pre-coated ELISA kits are guaranteed to have consistently high sensitivity, but other ELISA formats can encounter errors that reduce their sensitivity. Use this troubleshooting guide to avoid common sources of reduced ELISA sensitivity:

Problem Cause Solution
Incorrect assay type Assay format inherently not sensitive enough Switch to a more sensitive detection system
Switch to a more sensitive assay type
Use Boster high-sensitivity ELISA kits
Incompatible sample type Use sample types (e.g. serum vs. cell extract) verified to work as positive control
Include a positive control in your experiment
Reagent/sample problems Insufficient target Reduce dilution factor or concentrate sample
Inactive substrate Run a control to verify reporter enzyme reacts with substrate
Insufficient substrate Increase concentration of substrate
Mixing or substituting reagents from different kits Avoid mixing components from different kits
Plate problems Improper storage of ELISA kit Follow manufacturer storage recommendations
Poor target adsorption to wells Covalently link target to wells

Keywords: ELISA, Troubleshooting, Low Sensitivity, Low Detection Limit

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