Use of MAOS chromogenic substrate and enzyme preparation in ELISA experiment
In the enzyme-linked immunosorbent assay, the core detection reagents are MAOS, TOOS and other chromogenic substrates and related enzyme preparations. Enzymes are used as catalysts for the color reaction, and the chromogenic substrates are used as color indicators after the reaction. Through the color change, that is, the absorbance change, the purpose of detection can be achieved.
At present, the color-developing substrates TOOS, MAOS and various enzyme preparations are mostly needed in the automatic biochemical analyzer. The automatic biochemical analyzer is an instrument used to detect chemical components in human serum. It is mainly used to detect liver function, blood sugar, blood lipids, kidney function, and myocardium.
MAOS chromogenic substrate and enzyme preparation in ELISA experiment
With the popularity of biochemical analyzers, there are more and more problems in various aspects in use. In clinical testing, experimenters often mix different batches of reagents together. The purpose of this is: First, to save costs , The second is for the convenience of illustration. Obviously, no matter what kind of automatic biochemical analyzer manufacturer and which reagent, because of the different batch numbers of automatic biochemical analyzer reagents, there are the following differences:
1. The manufacturing time is different
2. The activity of the tool enzyme in the reagent is different
3. The concentration of the substrate that can produce hydrolysis will vary with time.
Especially for enzyme preparations, the potency of some enzyme preparations varies from batch to batch, and the activity varies with different storage times. Chromogenic substrates like TOOS may have small inter-batch differences and are not easy to cause influence, but TOOS after opening will slowly oxidize after contact with air, and should be used immediately.
Sometimes it takes a long time for the reagents of the automatic biochemical analyzer to open the bottles, which may cause bacteria to grow. Even if some reagents have preservatives, preservatives have limitations. In fact, most reagents do not contain antifungal ingredients. Therefore, in order not to affect the accuracy of clinical test results, reagent products such as chromogenic substrates and enzyme preparations are best to be used immediately, and it is generally not recommended to mix reagents of different batch numbers.
There is another point. Enzyme preparation manufacturers generally ship products under refrigeration and low temperature. If they are purchased, they need to be stored in refrigerator if they are not used immediately. Enzymes are protein preparations and avoid repeated freezing and thawing. Desheng's enzyme preparations are shipped refrigerated at 0-4 degrees, and chromogen substrates such as MAOS and TOOS are sealed and stored in opaque bottles.
When it comes to the chemiluminescent reagent Luminol, it is not unfamiliar to everyone and is commonly used in the field of criminal investigation to detect blood stains. Researchers have found that the principle of luminol luminescence can be used to detect specific substances in saliva, such as proteins, DNA, and related indicators of microorganisms, in order to determine physical conditions.