Acridine ester (NSP-DMAE-NHS), yellow powder, CAS No.: 194357-64-7, is an important chemiluminescent reagent with high quantum yield，mild reaction conditions, good reproducibility, and high chemiluminescence efficiency。NSP-DMAE-NHS is widely used in the fields of inorganic organic compounds, environmental monitoring, biological and pharmaceutical analysis, etc. In addition, it is also widely used for sensitive detection and diagnosis of many types of diseases.

In terms of in vitro diagnostics, acridinium ester(NSP-DMAE-NHS) compounds are well suitable for labeling DNA strands to make chemiluminescent DNA probes. This article will introduce a method of NSP-DMAE-NHS labeled nucleic acids.

Nucleic acid is the most important substance in all biological molecules, widely exists in all animal and plant cells, microorganisms, nucleic acid is divided into two categories: deoxyribonucleic acid (DNA) and ribonucleic acid (RNA). Modern medical research results show that the occurrence of many diseases, such as cancer and genetic diseases, is related to DNA mutation, and many infectious diseases are caused by viruses, pathogens or parasites in the environment. Therefore, the analysis of virus specific DNA sequence is conducive to the control of epidemic situation.

In nucleic acid hybridization analysis, the preparation of highly specific and sensitive labeled probes is the key to the success of nucleic acid hybridization analysis. Acridine ester(NSP-DMAE-NHS) derivatives can be directly labeled on nucleic acid probes without catalyst and the luminescence quantum yield is not affected. In addition, under certain conditions, the labeled acridinium ester(NSP-DMAE-NHS) on the unhybridized single-stranded DNA is hydrolytically destroyed, and only the double-stranded NSP-DMAE-NHS formed by the hybridization can produce chemiluminescence, and the entire hybridization process can be monitored without isolation.

This method of labeling nucleic acid with acridinium ester(NSP-DMAE-NHS)  comprises the following steps :

(1) protecting the 5'th terminal and the 3'the terminal of a DNA probe;

(2)  labeling acridinium ester: enabling 25mm NHS acridinium ester to dissolve in DMSO, preparing 1M of an HEPES buffer solution (PH=8.0), according to the condition that the molar ratio of the nucleic acid probe to the NHS acridinium ester is 1 to 5, performing adding to the HEPES buffer solution, and performing a reaction at 37 DEGC for 1h;

(3) performing HPLC purification. The method has the beneficial effects that according to an ordinary method, one acridinium ester can be labeled at the 5' terminal of the nucleic acid probe.

The method creatively labels acridinium ester(NSP-DMAE-NHS) at two ends, and further improving the sensitivity of detection.

Related links:NSP-DMAE-NHS

Reference: Method of  NSP-DMAE-NHS labeled nucleic acid CN109852669A