ELISA failure may be caused by many factors. Many unexpected results can be avoided if reading and fully understanding the manual before experiment. You can refer to the tips as following to identify your assay failure.
1. Check the expiration date of the kit, ensure that all the reagents were stored as suggested in the manual.
2. Check for signs of instability or deterioration in reagent solutions, such as precipitation or discoloration and so on.
3. The substrate reagent should be colorless.
4. Use clean plastic disposable pipettes, tips, and containers for reagent preparation and storage. Avoid cross-contamination of
reagents by changing pipette tips between addition of each standard, sample and reagent.
5. Ensure that specified incubation times and temperatures have been guaranteed.
6. Ensure that no substitution of kit reagents has occurred.
7. It is recommended that samples and standards should be run in duplicate to improve accuracy.