BM Chemiluminescence Western Blotting Kit (Mouse/Rabbit)

Uses

BM Chemiluminescence Western Blotting Kit (Mouse/Rabbit) is used to detect proteins with peroxidase-labeled secondary antibodies and the chemiluminescent substrate luminol on western blots. The chemiluminescence system is specifically suited for western and dot blot applications where high sensitivity is required. Chemiluminescence detection of proteins is 1 to 3 orders of magnitude more sensitive than colorimetric methods. Compared with radioactive detection of blotted antigens, the chemiluminescence image is at least as sensitive after much shorter exposure times and without the risk of working with radioactivity.

General Description

Detection of membrane-bound molecules is carried out by a variety of procedures. In protein analysis, the molecules of interest are usually detected with specific first antibodies, enzyme-labeled secondary reagents (antibody- or streptavidin-conjugates) and a suitable chromogenic substrate. These detection systems, however, may be time-consuming or lacking sensitivity. In addition, colors tend to fade with time and exposure to light.
To overcome these problems, attempts have been made to use chemiluminescence for fast and sensitive detection of enzyme conjugates on blots. The BM Chemiluminescence Western Blotting System is designed around peroxidase-labeled secondary antibodies and the substrate luminol. In the presence of hydrogen peroxide (H2O2), horseradish peroxidase (POD, HRP) catalyzes the oxidation of diacylhydrazides such as luminol. An activated intermediate reaction product is formed, which decays to the ground state by emitting light. Strong enhancement of the light emission is produced by 4-iodophenol. This acts as a radical transmitter between the formed oxygen radical and luminol.

Raw materials

Preparation Products