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Do you know the reasons for the deterioration of ELISA kits?
Deterioration of ELISA kits is a common problem in ELISA experiments. So, what is the reason for the deterioration of ELISA kits? What affects the warranty of ELISA kits? What impact will it have once it deteriorates? Hengyuan Biology will provide a detailed analysis for everyone.
1、 The impact of methodology
The ELISA detection modes include the following: double antibody sandwich method, indirect method, double antigen sandwich method, IgM antibody capture method, and competitive inhibition method. Among them, the competitive inhibition method (used for HBeAb, HBcAb, etc.) has poor repeatability and difficult quality control due to factors such as uneven competition caused by operation time difference.
2、 Reagent factors
It is difficult to ensure that the quality of ELISA reagents from different batches is completely consistent during the production process. Even for projects that pass batch testing, the test results may vary. Therefore, it is necessary to choose and order long batch reagents and ensure storage conditions. Strictly implementing this standard can avoid the complex process of re establishing quality control systems and re evaluating reagents due to changes in reagent batch numbers, and can ensure the stability of results; For reagents with short shelf life and low usage rate, they should be packaged in small quantities, and the packaged portion should be taken each time to avoid repeated freezing and thawing that may cause reagent failure.
3、 Sample factors
Specimen interference factors include endogenous interference factors and exogenous interference factors. The former includes rheumatoid factors, complement, heterophilic antibodies, autoantibodies, lysozyme, etc. The latter includes specimen hemolysis, bacterial contamination, prolonged specimen storage time, incomplete specimen coagulation, and repeated freezing and thawing of frozen specimens.
4、 Operational factors
The operation steps of ELISA are complex, and improper operation can cause significant errors. Sample addition, incubation, washing, color development, and color comparison.
5、 Setting of gray area
Normally, ELISA qualitative experiments report results as "positive" and "negative", with a boundary between the two known as the "cut-off value" (CO value), which serves as the basis for reporting qualitative immunoassay results.
6、 Sample re examination
The manual ELISA detection process is complex and has many influencing factors. Even if indoor quality control is in place, differences in results may still occur due to differences between wells. Therefore, increasing the intensity of re examination is a reliable method to ensure the accuracy of results, such as re examination of suspicious, weakly positive specimens and rare patterns of HBsAg, HBeAg, HCV Ab, HIV Ab, TP Ab and other items.
7、 Common methods for expressing results frequently
1. Qualitative determination
2. Semi quantitative determination results are generally expressed in terms of titers.
3. Quantitative determination refers to using a known amount of standard substance for a series of dilutions and conducting ELISA analysis, drawing a standard curve, and expressing the results in absolute quantities or units. In ELISA quantitative detection, each reaction plate must be plotted with a corresponding standard curve.
Hengyuan Biotechnology is a company that produces and sells reagents for scientific research and development, covering basic innovation fields such as chemistry, analytical chemistry, life sciences, and materials science. With the help and support of a large number of scientific research users, Hengyuan Biotechnology has made unremitting efforts to provide customers with a stable platform with excellent product quality and good service attitude, becoming a reliable long-term partner for customers.
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