2 - ELISA Weak/No Signal? Causes & Solutions for Reliable Results

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Possible Causes & Recommended Actions

Reagent Mixing

Reagent Mixing

Do not mix reagents from different batches or different indicators for the same indicator.

Improper Reagent Storage

Improper Reagent Storage

Store according to the manuals.

Expired reagents

Expired reagents

Check the expiration date of the kit before use.

Missing or Incorrect Addition of Detection Antibodies/Enzyme Conjugates

Missing or Incorrect Addition of Detection Antibodies/Enzyme Conjugates

Prepare and add according to the manuals, check the liquid level and remaining amount at each step, and verify the label information before adding samples.

Low Incubation Temperature or Short Incubation Time

Low Incubation Temperature or Short Incubation Time

Follow the instructions for operation. The ELISA kits from Linkage Biosciences are incubated at room temperature (25°C±3°C). The one-step ELISA kits are incubated at room temperature/37°C.

Incorrect Order of Reagent Addition

Incorrect Order of Reagent Addition

Add samples according to the manuals, and verify the reagent labels at each step.

Contamination of Detection Antibodies/Enzyme Conjugates/Buffers

Contamination of Detection Antibodies/Enzyme Conjugates/Buffers

Prepare new dilutions of detection antibodies/enzyme conjugates/buffers.

Inaccurate Pipette, Incorrect Sample Addition

Inaccurate Pipette, Incorrect Sample Addition

Ensure sufficient dissolution of microspheres in each well of the one-step experiments and control the time interval for sample loading in each well as much as possible.

Insufficient Dissolution of Microspheres in One-step Experiments

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Ensure sufficient dissolution of microspheres in each well of the one-step experiments and control the time interval for sample loading in each well as much as possible.
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Insufficient Washing or Thorough Washing in One-Step Experiments

Insufficient Washing or Thorough Washing in One-Step Experiments

Use a new batch of washing solution in the one-step experiments, ensure clean and residue-free wells after washing; extend the interval for plate washing or increase the number of washes.

Moisture Dissolution of Microspheres in One-Step Experiments

Moisture Dissolution of Microspheres in One-Step Experiments

Avoid exposing microspheres to high humidity environments.

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