Chemiluminescence reagent acridine ester labeling

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Labeling principle of chemiluminescence substrate acridine ester in diagnostic reagent in chemiluminescence immunoassay, we use acridine ester to label the antibody protein first, and then we can detect the substance to be tested after the immune reaction. Therefore, how to label the antibody is very key. Acridine salts and related compounds have been widely proved to be very useful chemiluminescence markers. Their stability, labeling specificity and detection sensitivity are better than radioisotopes.

Acridine NHS ester, namely acridine succinimide ester (provided by this kit), can react with the primary amino group of protein. Under alkaline conditions, NHS is replaced as a leaving group, and protein forms a stable amide bond with acridine ester. After the reaction is completed, the excess acridine salt is removed through the desalination column. In the presence of alkaline hydrogen peroxide, acridine labeled proteins can emit light without enzyme catalysis. Therefore, the addition of excitation solution causes the reaction system to immediately release about 430nm photons. The concentration of protein can be detected by counting the number of photons with a standard photometer luminometer. Because the luminescence process is very short (the whole process is completed in 2 seconds), the sample must be placed directly in front of the photon detector inside the photometer. Proteins, peptides, antibodies and nucleic acids can be labeled with acridine ester.

Chemiluminescence reagent it should be noted here that when dissolving acridine succinimide ester, the DMF used must be strictly anhydrous to prevent the hydrolysis of acridine ester. When not in use, it should be dried and sealed at - 20 ° C for storage. Aiming at the traditional acridine ester, its structure was modified to increase the steric hindrance and enhance the hydrolysis resistance. For example, it is stable in Pb buffer with pH 7.0 at room temperature. If the acridine carboxylic acid does not contain - NHS, it needs to add condensing agent EDCI, etc. in order to have coupling reaction with the protein containing amino group. Acridine hydrazide contains free amino groups. The terminal of acridine hydrazide is suitable for direct coupling of polysaccharide nucleic acids or proteins containing aldehyde groups.

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