In chemiluminescence immunoassays, we need to label the antibody protein with acridinium esters before the test object can be detected after the immune response. Therefore, how to label the antibody is very critical. Acridinium ester compounds have been widely proven to be very important. Useful chemiluminescent labels, their stability, labeling specificity and detection sensitivity are superior to radioisotopes.
Principle of acridine ester luminescent labeling:
Under alkaline conditions, NHS is substituted as a leaving group, and the protein forms a stable amide bond with the acridine ester. After the reaction is completed, the excess acridine salt is removed through a desalting column. In the presence of alkaline hydrogen peroxide, acridine-labeled proteins can emit light on their own without enzymatic catalysis.
The addition of the excitation solution causes the reaction system to immediately release photons of about 430 nm. The protein concentration can be detected by counting the number of photons with a standard luminometer. Because this light-emitting process is very short (the whole process is completed within 2 seconds), the sample must be placed directly in front of the photon detector inside the photometer. Proteins, peptides, antibodies, and nucleic acids can all be labeled with acridinium esters.
Precautions for acridine ester luminescent label
There are many types of acridine esters. Common acridine esters, acridine sulfonamides, acridine sulfonates, etc. are all acridine ester compounds. It should be noted that when dissolving the acridine succinimide ester, the DMF used must be strictly anhydrous to prevent the acridine ester from being hydrolyzed. When not in use, store it in a dry seal at -20°C.
If it is acridine carboxylic acid that does not contain -NHS, it is necessary to add a condensing agent EDCI, etc., in order to have a coupling reaction with an amine-containing protein. Acridine hydrazide contains free amino groups, so it is more suitable for direct coupling to polysaccharides, nucleic acids or proteins containing aldehyde groups.
Acridine ester luminescence advantage
1. For traditional acridine esters, the structure has been modified to increase steric hindrance and enhance hydrolysis resistance. For example, at room temperature, it is stable in a PB buffer with a pH of 7.0.
2. The light release is fast and concentrated, the luminous efficiency is high, the luminous intensity is high, it is easy to connect with the protein and the photon yield does not decrease after the connection.
3. There are few interference factors in the luminescence reaction, and the label is stable (can be stored for several months at 2-8 ℃)
The acridinium esters developed and produced by Desheng Technology belong to independent research and development and production, and a lot of energy has been invested in the research and development of acridinium esters, so the quality control is more stringent. At present, the company's products have been sold to more than 100 countries around the world, and more than 99% of customers have received good reviews for repurchase. The product quality is excellent and the price is favorable. If you are interested in understanding, you can call for consultation. Desheng welcomes your calls.
