[ELISA kit] How to reduce the experimental error probability? In the process of operation, in addition to the need to be more careful, there are some places that need to be paid attention to. Shanghai Hengyuan Biological Co., Ltd. has summarized nine aspects for you:
Aspect 1: Inspection
Should have the basic quality and practical experience of doing laboratory operations. Proficiency in the operation of experimental instruments and instruments, with the ability to summarize and analyze problems, and to solve the accidents in the experiment in a timely manner.
Aspect 2:
Use a proofread micropipette to eliminate natural errors. Whether the pipette is accurate or not is especially important for quantitative detection.
Aspect 3: Read the instructions carefully
Standardize operations in strict accordance with the requirements of the manual. Reagents of different batches cannot be mixed. Where it is worth improving, trial and error can be improved after it is established.
Aspect 4: Wash thoroughly
The washing plate is not thorough, and the background color of the enzyme conjugate will show a false positive. The washing liquid should be fresh, and the existing washing liquid will have a background increase, and false positives may also occur.
Aspect 5: Strict control of reaction time
The reaction time is too long, the enzyme is inactivated; the reaction time is too short, the enzyme conjugate cannot be fully combined with the microbial antigen antibody in the serum, and the structure of the product is loose and not firm, and it is easy to wash off, which may cause false negative.
Aspect six
Note [ELISA kit] During the shelf life, reagents that have passed the shelf life cannot be used.
Aspect 7: Applicability of the evaluation reagent
Whether the reagent is stable or not is important for the accuracy. Before the reagent is activated, the yin and yang control and the sample repeated comparison test should be carried out, and the reagent can be used after determining that the reagent meets the requirements.
Aspect 8: Strictly master the color development time
The color development time is too short. After the termination reaction is terminated, the amount of the substrate conjugate is too small, and false negative is likely to occur. Color development after the time required for color development should be judged as a false positive, which may be related to the reagent itself. Immediately after the coloring agent is added, the color is developed, and no positive is reported. This may be the result of background color development.
Aspect 9: Adding samples should be accurate and fast
If the sample is not accurate, the amount of enzyme production cannot be determined, directly affecting the color development result. The color depth and the determination of the A value are related to the amount of the developer and the stop solution added, so the sample should be used with caution. If the sample is loaded within a certain period of time, if the sample is sluggish, the error will occur and the reagent will be exposed for a long time. Especially when the room temperature is too high, the shelf life will be shortened or even invalid.
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