How to label nucleic acid with acridine ester luminescent reagent
Acridinium esters are important luminescent markers in chemiluminescence immunoassays. They can be used for the detection of proteins, antibodies and nucleic acids. These substances can be labeled for the detection of proteins, antibodies and nucleic acids. Among them, there is a certain difference between the labeling of nucleic acid and the labeling of protein. This article introduces how to label nucleic acid with acridinium ester.
Types of acridinium esters for labeling nucleic acids
Take Desheng’s acridine ester luminescence reagent as an example. There are currently 6 types of acridinium esters developed. Among them, the N-hydroxysuccinimide group (NHS ester) is usually used for protein labeling. Acridine hydrazide NSP-SA-ADH can be used for nucleic acid. This acridinium ester end is suitable for direct coupling to label the aldehyde group in the nucleic acid. Of course, it can also be labeled with aldehyde group polysaccharides, nucleic acids or proteins.
Significance of acridine ester labeling nucleic acid
Labeling DNA strands with acridine ester chemiluminescent reagents can produce chemiluminescent DNA probes for in vitro diagnosis. Nucleic acids include DNA and RNA. Many diseases such as cancer and genetic diseases are related to DNA mutations. In addition, the analysis of virus specific sequence DNA is conducive to the control of the epidemic.
Method for labeling nucleic acid with acridinium ester
The DNA probe made of acridinium ester-labeled nucleic acid has strong specificity and high sensitivity, which is the key to successful nucleic acid hybridization analysis. Firstly, the 5'end and 3'end of the DNA probe were protected respectively; then the acridinium ester labeling was carried out, and finally the DNA was purified and separated by HPLC. Among them, the acridinium ester labeling is the most important. Dissolve 25mM acridinium ester in dimethyl sulfoxide (DMSO), configure 1M HEPES buffer (PH=8.0), according to the molar ratio of nucleic acid probe: acridinium ester=1:5 Add to HEPES buffer and react at 37°C for 1h.
The above method creatively labeled acridine esters on both ends of the DNA, which further improved the sensitivity of detection. Acridine esters have the advantages of mild reaction conditions, good reproducibility, high luminous efficiency, and strong luminous intensity. After the nucleic acid is labeled with acridine ester, a chemiluminescence reaction is performed without a catalyst and the luminescence quantum yield is not affected.
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