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4.Enzyme marker inactivation or substrate failureMix the enzyme conjugate and substrate and check and judge by rapid color development Problem description: Low readingsPossible causes and corresponding countermeasures: 1.The microplate reader is not set up correctlyCheck wavelength and filter settings on the microplate reader/Turn on the ELISA reader in advance to preheat Problem description: Large coefficient of variationPossible causes and corresponding countermeasures: 1.Incorrect addition of liquidCheck the filling status Problem description: High background valuePossible causes and corresponding countermeasures: 1.The working concentration of the detection antibody is too highUse the recommended dilution factor 2.Incomplete washing of ELISA plateEnsure that each step is cleaned completely

doi: 10.3389/fcell.2022.808859 16 GarofanoFSchmidt-WolfIGH