Luminol luminol is an earlier chemiluminescence substrate, such as luminol luminescent fluid, ECL, isoluminol reagent, ABEI, AHEI, ITCI, etc. are all based on luminol or luminol derivatives Chemiluminescence immunoassay reagents. Among them, ECL luminescent reagent is a kind of reagent with more applications based on luminol.

Luminol luminol is a traditional luminescent substrate. The ECL chemiluminescence detection reagent prepared therefrom has better luminescence performance than luminol. It belongs to a new generation of enhanced chemiluminescence substrate reagents and can be coupled with secondary antibodies. The POD or horseradish peroxidase (Horseradish Peroxidase, HRP) catalyzes a chemical reaction to emit fluorescence. The result can be displayed by X-ray film pressing and other imaging techniques or detected by Luminometer.

The ECL ultra-sensitive luminescent solution made from luminol decays slowly, and the fluorescence will hardly be weakened within 15 minutes, and the non-specific background generated when the detection is less interference is very low, which can reduce the amount of antibodies and samples to be tested. Western blot of PVDF membrane and NC membrane (nitrocellulose membrane) and other blots are luminous.

The protein or nucleic acid substrate can be directly or indirectly detected by ECL after being labeled with horseradish peroxidase HRP. It has extremely high sensitivity and high signal-to-noise ratio. It can detect 10-100 fg antigen (#P1010); it emits fast and belongs to Long-time glow type, fluorescence can make X-ray film photosensitive for more than 12 hours, especially suitable for trace protein or nucleic acid detection. At the same time, the reagent allows the use of higher antibody dilution ratios (1:2000-1:10000), which greatly saves antibodies.

There are a few points to pay attention to when using ECL luminescent liquid:

1. When using ECL, the previous steps can be carried out in sunlight, but the sensitivity may be slightly reduced when exposed to strong light for too long. Wearing gloves can avoid leaving fingerprints on the film.

2. If the exposure is too long or the protein is too much, the background will deepen and the band strength will lose the linear relationship, while the underexposure will blur the band.

3. The strips on the membrane glow after incubating for about 1 minute in the luminescent working solution. The strong band luminescence is visible to the naked eye in the darkroom, and the low-abundance protein band luminescence is weak or even invisible to the naked eye, but it can be exposed to X-ray film. The light emission time of the strip cannot be judged by visual observation alone.

4. Too high antibody concentration will cause high background or no bands, which will lead to failure!

ECL luminescent fluid is mainly made of luminol, isoluminol or isoluminol derivatives, sodium hydroxide and hydrogen peroxide. Desheng produces luminol and isoluminol raw materials, which need to be matched Lye, hydrogen peroxide, and peroxidase can complete the chemiluminescence experiment.