Acridine ester (nsp-dmae-nhs), yellow powder, CAS No.: 194357-64-7, is an important chemiluminescence reagent. Its chemiluminescence quantum yield is higher than that of luminol. Moreover, the labeling conditions of acridine ester are mild, the labeling rate is high, and the labeling does not affect the separation.
In addition, the chemiluminescence process of acridine ester is rapid with low background, and can emit light in the presence of sodium hydroxide and hydrogen peroxide. In addition, acridine ester chemiluminescence reagents have good stability and are easy to store.
In addition to the application in immunoassay, acridine ester can also be used to label oligonucleotide fragment probes for gene detection or microbial detection. Acridine ester compounds are suitable for labeling DNA strand to make chemiluminescence DNA probes.
Modern medical research results show that many diseases such as cancer and genetic diseases are related to DNA mutation, and many infectious diseases are caused by viruses, bacteria or parasites in the environment. The analysis of specific sequence of virus DNA is conducive to the control of epidemic situation. The detection of DNA sequence and base mutation in its strand is of great significance in gene screening, early diagnosis and treatment of genetic diseases, and detection of pathogenic genes.
In nucleic acid hybridization analysis, the preparation of specific and sensitive labeled probe is the key to the success of nucleic acid hybridization analysis. Acridine ester derivatives can be directly labeled on the nucleic acid probe without catalyst and the luminescence quantum yield is not affected. In addition, under certain conditions, the acridine ester labeled on the unhybrid single strand DNA is hydrolyzed and destroyed, and only the double strand formed by hybridization is protected Only acridine ester can produce chemiluminescence, and the whole hybridization process can be monitored without separation.
The invention relates to a method for labeling nucleic acid with acridine ester, which comprises the following steps:
(1) The 5 'and 3' ends of DNA probe were protected;
(2) Acridine ester labeling: 25 mm NHS acridine ester was dissolved in DMSO, and 1 m HEPES buffer (pH = 8.0) was prepared. According to the molar ratio of nucleic acid probe: NHS acridine ester = 1:5, it was added into HEPES buffer and reacted at 37 ℃ for 1 h;
(3) Purified by HPLC. In the step (1), the protection of 5 'and 3' ends of DNA probe specifically includes: using s-ethyl trifluorothioacetate to protect 5 'end-nh 2; using s-ethyl trifluorothioacetate to protect 3' end-nh 2.
Desheng technology has been researching and developing chemiluminescence reagents for 12 years. After successful development, it has been put into the market and won the general recognition of customers. There are six different groups of acridine esters. Besides acridine esters, there are luminol and isoluminol. Acridine ester series have high luminous efficiency and belong to direct luminescence.
