Comparison of chemiluminescence and fluorescent substrates of HRP in enzyme-linked immunoassay

The most commonly used enzyme preparation in enzyme-linked immunoassay is HRP horseradish peroxidase. There are three types of substrates in biochemical detection, and there are mainly three types, namely chromogen substrates such as TOOS, MAOS, and fluorescent substrates. Substrates, chemiluminescent substrates such as luminol. Among them, chemiluminescence substrates are also used in chemiluminescence analysis, which is becoming more and more popular.

Whether it is a chromogen substrate, a fluorescent substrate or a chemiluminescence substrate, it is used as a labeling and detection method, which plays a key role in the accuracy and sensitivity of the detection result. The most popular one is chemiluminescence immunity. analysis.

Luminol HRP Chemiluminescence Substrate

The development of different substrates in biochemical testing:

In terms of the development time of the detection method, the fluorescent substrate is the product of peroxidase developed after the chromogen substrate, and the detection sensitivity is higher than that of the chromogen substrate. Several fluorescent materials of different colors can be used at the same time. , But the equipment is expensive, the detection cost is high, and it is easily interfered by autofluorescence generated by cells or other molecules, and it is sensitive to light and unstable during observation and storage. After fluorescent substrates, the discovery of luminol-like chemiluminescent substrates brought horseradish peroxidase HRP substrates into a rapid development stage, and various chemiluminescent substrates greatly enriched horseradish peroxidase Applications.

The difference between chemiluminescence and fluorescence:

Chemiluminescence is different from fluorescence. It means that the chemical energy generated in the process of chemical reaction is absorbed to stimulate molecules and emit light. Fluorescence is the light emitted by absorbing light energy to excite molecules. This is chemiluminescence and The difference in fluorescence lies in the different forms of excitation energy that form excited molecules between the two, one is chemical energy and the other is light energy. Chemiluminescence detection has high sensitivity and low background, so it is widely used.

Currently common chemiluminescent substrates, including luminols, peroxyoxalates, and acridine esters, are several chemiluminescent reagents commonly used in actual chemical analysis and determination, and are used in drug analysis, environmental monitoring, etc. The field also has important applications. The above-mentioned chemiluminescence reagents are specially produced by Desheng, but it should be noted that acridine esters can emit light directly and do not require enzyme catalysis.