Precision Detection and Experimental Exploration of the Chemiluminescence Efficiency of Acridine Esters

In the vast field of in vitro diagnosis, chemiluminescence immunoassay has attracted much attention for its high sensitivity and accuracy. Among them, acridine ester has become a leader in chemiluminescence reagents due to its high detection sensitivity and quantum yield. The luminescent reaction of acridine ester does not require enzyme catalysis, which gives it a special advantage in the field of biological detection. So, how to accurately detect the luminescence efficiency of acridine ester? Next, we will delve deeper into its detection methods.

The basic principle of acridine ester chemiluminescence efficiency detection

The core of chemiluminescence efficiency detection lies in measuring the number of photons per second of acridine ester excited. This process is usually completed through a photometer. By measuring the luminescence intensity of different concentrations and volumes of acridine ester solutions, we can obtain specific values of their luminescence efficiency.

Acridine ester powder

Experimental preparation and operation

1. Reagent preparation

Before starting the experiment, we need to prepare a series of high-quality reagents, including dimethylformamide DMF (purity>99%), hydrogen peroxide, nitric acid, sodium hydroxide analytical pure, cetyltrimethylammonium chloride (Ctac), Tween-20, TritonX-100, etc. At the same time, it is necessary to prepare 96 well plates and monoclonal antibodies against human TSH.

2. Luminescent start and measurement

Take an appropriate amount of DMAE-NHS solution into a 96 well plate (be careful to use a black opaque well plate to avoid light interference). Then, add the luminescent starting reagent and measure the change in luminescent count over time on the luminescent instrument. For NSP-DMAE-NHS or other types of acridine esters, similar measurement methods should also be used.

Experimental optimization and exploration

1. Optimization of luminescent starting reagents

By changing the concentration and volume of each component (H2O2, HNO3, and NaOH) in the luminescent activation reagent, we can measure the luminescence intensity corresponding to different concentration volumes. This step helps us analyze the effect of luminescent starting reagents on the luminescence intensity of DMAE-NHS, thereby optimizing experimental conditions.

2. Exploration of surfactants

Add surfactants Ctac and Tween-20,TritonX-100, We can measure changes in luminous intensity. This step helps us understand the effect of surfactants on chemiluminescence. However, it should be noted that although some surfactants can enhance chemiluminescence, surfactants such as Ctac may cause an increase in background, so they are not suitable as enhancers for acridine ester DMAE-NHS chemiluminescence.

The characteristics and advantages of acridine ester chemiluminescence

Compared with luminol and other glow type chemiluminescent reagents, acridine ester belongs to the category of flash type chemiluminescent reagents. Its luminous time is brief (usually only 2-4 seconds), so it is necessary to use a photometer for real-time detection. However, it is precisely this flashing characteristic that gives acridine esters a special advantage in the field of biological detection. Its high sensitivity and quantum yield enable it to be used for detecting low concentrations of biomolecules, providing strong support for the detection of physical health.

Product packaging

As a company dedicated to the research and development of biological detection reagents, Desheng has successfully developed various acridine ester based chemiluminescent reagents. In addition to DMAE-NHS, Desheng has also developed five other acridine ester reagents that can be used for labeling different types of proteins, antibodies, and nucleic acids. The successful development and application of these reagents will undoubtedly bring more convenience and possibilities to the field of biological detection. If you have any purchasing needs in the near future, please click on the Desheng official website to learn more details or contact me!