Common errors in protease inhibitor testing include:
Expired reagents or mixed components of different reagent kits: Using expired reagent kits or mixing components of different reagent kits may result in inaccurate experimental results. It should be ensured that the reagent kit has not expired and all components belong to the corresponding reagent kit.
Incorrect or missed addition of reagents, substrates, and color reagents: Negligence during operation may result in incomplete color development steps, affecting the accuracy of experimental results. It is necessary to strictly follow the instructions and check whether the liquid level is consistent to reduce leakage.
Enzyme label contamination and inactivation: If the container containing the enzyme label contains enzyme inhibitors (such as sodium azide), or if the enzyme label is contaminated during plate washing and sample addition, it may cause the enzyme to lose its ability to catalyze the color development of the color reagent. Ensure that the container does not contain enzyme inhibitors and that the washing solution container has been cleaned thoroughly.
There is a problem with distilled water: If the distilled water used to prepare the washing solution does not meet the requirements or is contaminated, it may affect the experimental results. It should be ensured that the distilled water meets the requirements and is not contaminated.
The incubation time and temperature did not meet the requirements: The temperature and incubation time of the reaction plate when placed in the incubator should be strictly controlled to ensure that the reaction proceeds fully. The incubation temperature should be controlled at 37-38 ℃, and the incubation time should be strictly operated according to the instructions.
Excessive washing of plates or inadequate dilution ratio of concentrated washing solution: Improper parameter settings during the washing process may affect the experimental results. It is necessary to strictly dilute and concentrate the washing solution and wash the plate according to the instructions, reduce the washing impact force, and keep the washing solution for the required time and number of washes according to the instructions.
These errors may affect the accuracy and reliability of protease inhibitor assays, so experimenters should pay attention to operational details and ensure that experimental conditions meet requirements to obtain accurate results.
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