CAS 521-31-3 Luminol In Protein WB Experiment Luminol Luminescence Reaction

Application Of Luminol CAS521-31-3 In Protein WB Experiment Luminol Luminescence Reaction

The chemiluminescence reagent luminol, the full name of 3-aminophthalic hydrazide, and the common name of luminol, is usually used for enzymatic chemiluminescence experiments and detection of trace blood stains in criminal investigations. Because of its simple structure, easy synthesis, and high luminous quantum efficiency, it is now also used in western blot for western blotting.

Basic information

Western blotting is the transfer of protein samples separated by PAGE (polyacrylamide gel electrophoresis) to a solid carrier NC membrane (nitrocellulose membrane) or PVDF membrane (polyvinylidene fluoride membrane). The non-covalent bond form adsorbs proteins, and can keep the types of peptides separated by electrophoresis and their biological activities unchanged. Use the protein or polypeptide on the solid carrier as the antigen to react with the corresponding antibody, and then react with the enzyme or isotope-labeled second antibody, and then undergo color development or autoradiography to detect the specific purpose of electrophoretic separation The protein component of gene expression. This technique is also widely used to detect the expression of protein levels.

Luminol luminescence reaction

Luminol's luminescence reaction is the last step in the Western Blot blotting experiment of the entire protein. Add 1:1 solution A and solution B to cover the pvdf film, and you can develop color or luminescence to observe the experimental results. The commonly used luminescent liquids A and B are luminol and hydrogen peroxide, respectively.

Since the reaction of luminol catalyzed by hydrogen peroxide to oxidize and emit light requires the participation of peroxidase, it belongs to enzymatic chemiluminescence and needs to maintain the pH value of the reaction. The homogenization buffer in the experiment is usually made of our Tris-HCL TBS buffer, protein transfer buffer also use Tris plus SDS, glycine, methanol buffer system, if it is higher than 20kd protein, you can use CAPS buffer, and use Tricine instead of glycine.

Luminol is usually made into ready-to-use ECL luminescent liquid on the market. Luminol produced by Desheng is a powder reagent. Because the luminescent reagent is sensitive to light and temperature, the powder reagent is more conducive to storage. It can be used, and different concentrations can be configured according to different experiments. There is no need to purchase multiple concentrations of reagents to reduce waste.