BM Chemiluminescence Western Blotting Kit (Mouse/Rabbit)

100

Roche

wet ice

2-8℃

1 kit containing 4 components.

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.

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.

Each lot is function tested on a dot blot.
The BM Chemiluminescence Western blotting System is designed for peroxidase-labeled secondary antibodies and the substrate luminol. In the presence of hydrogen peroxide (H202), horseradish peroxidase (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.

Working solution: Preparation of additional reagents and solutions
The table inside the PI describes the preparation of working solutions. For reproducible results equilibrate all solutions to 20 to 25 ℃ before use.


Storage conditions (working solution):

• 1% Blocking solution: -15 to -25 ℃
• 0.5% Blocking solution: -15 to -25 ℃
• POD-labeled secondary antibody stock solution: 12 month at 2 to 8 ℃
• POD-labeled secondary antibody working solution: prepare always fresh
• Detection solution: 1 week at 2 to 8 ℃

Reconstitute lyophilizate in 100 μl double-dist. water. This solution is stable for 12 months at 2 to 8 ℃.

仅用于生命科学研究。不可用于诊断。

组份不可单独销售
Luminol Substrate Solution with enhancer
NaBO₃ Starting Solution
Blocking Reagent
Anti-mouse IgG-POD antibody, Fab fragments/Anti-rabbit IgG-POD antibody

GHS07

Warning

H317

P261 - P272 - P280 - P333 + P313 - P362 + P364 - P501

Skin Sens. 1

12 - Non Combustible Liquids

WGK 2