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.