Food Allergen ELISA

Food Allergen ELISA Kit - Troubleshooting

  • High C.V. value
  • High background
  • Color development is poor, or color cannot be achieved even after adding chromogenic substrate solution.
  • Developed color but can’t be measured

1. High C.V. value

Possible reasons :
Mixing occurred during separate injection
When separately injecting the solution to the plate, the solution may have splattered and mixed with the neighboring well.

The plate bottom is dirty.
The back of the plate may be contaminated with washing solution, or absorbance was not correctly measured. Wipe the bottom of the plate with a paper towel, etc., and measure absorbance again.

Inadequate washing operation
Check the amount of washing, washing frequency, washing method, etc. Washing with pipette may dry the well, making absorbance higher. Use of an ELISA washing dispenser is recommended.
Also be sure not leave any reactant or washing solution in the well when washing. Carry out calibration when using a plate washer, and make sure the washing solution sufficiently fills each well.

Error during separated injection
Check pipetting operation, etc.
Plate drying
Promptly inject the next reagent separately.

2. High background

Possible reasons :
Insufficient washing operation
Check the amount of washing, washing frequency, washing method, etc. Washing with a pipette may dry the well, making absorbance higher. Use of a ELISA washing dispenser is recommended.
Also be sure not leave any reactant or washing solution in the well when washing. Carry out calibration when using a plate washer, and make sure the washing solution is sufficiently fills each well.

Reaction temperature is high
Be sure to return the product to room temperature before use. And, maintain the proper room temperature when carrying out reaction.
Reaction time is long.
Be sure to maintain an accurate reaction time.

Plate drying
Promptly inject the next agent separately.
Contamination of the washing reagent and the micropipette
Soak all equipment, such as the washing bottle, in an alkali detergent, and wash with an ultrasonic cleaner. Contamination from the micropipette can be avoided by using pipette chip with a filter.

3. Color development is poor, or colored cannot be achieved even after adding chromogenic substrate solution.

Possible reasons :
Reaction temperature is low
Be sure to return the product to room temperature before use. And, maintain the proper room temperature when carrying out reaction.
Reaction time is short
Be sure to maintain an accurate reaction time.
Degradation due to condensation of solid-phase antibodies
After returning the plate to room temperature, open the aluminum bag.
Plate drying
Promptly inject the next agent separately.
Insufficient amount of solution.
Check pipetting operation, etc.
The wrong enzyme-labeled antibody solution has been added
Check if any other solution has been added, and that operation has been carried out in the correct order.
Improper storage of the kit
Store the kit between 2-8℃. Return the product to room temperature before use.
Absorbance measurement was not carried out promptly
Carry out absorbance measurement within 30 minutes after enzymatic reaction has been stopped.

4. Developed color but can’t be measured

Possible reasons :
Plate reader error
The plate reader may not be operating normally. Check for any abnormality in the power source or lamp, and that the measurement wavelength is correct.

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