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Chemiluminescent Reagents for Western Blotting
Chemiluminescence is a phenomenon in which the energy produced by a chemical reaction is emitted in the form of light; chemiluminescent reagents use this phenomenon for the detection of proteins and nucleic acids1,2) as well as for in vivo and cellular imaging.3) Many types of chemiluminescence reagents exist, all using different types of enzyme and/or substrate. In Western blotting, for example, one of the most common detection methods employs horseradish peroxidase (HRP) to catalyze the oxidation of luminol, releasing light.1)
Chemiluminescence HRP substrate solution kit is a reagent that allows chemiluminescent detection of HRP-labelled probes bound to proteins on membranes. The kit consists of two components and should be mixed at a ratio of 1:1 just before use. Although the detection sensitivity depends on the experimental conditions, highly sensitive detection is possible like in the below figure.
Figure. Membrane luminesced with chemiluminescence HRP substrate solution kit
HeLa Cell Lysate: 2.5 - 0.039 μg/lane (2x step dilution), Membrane: PVDF membrane, Blocking Buffer: 1% BSA/TBS-T, Primary Antibody: Anti-GAPDH (Mouse IgG),
Secondary Antibody: Goat anti-Mouse IgG HRP, Detection: CCD Imager, Exposure Time: 120 seconds, photos were taken at TCI.
Related Products
- C4087
- Chemiluminescence HRP Substrate Solution Kit [for Western Blotting]
- W0024
- Western Blot Stripping Buffer [for Biochemical Research]
- G0407
- Goat Anti-Mouse IgG HRP Conjugate
- G0418
- Goat Anti-Rabbit IgG HRP Conjugate
- S0972
- Streptavidin HRP Conjugate
- D5909
- DAB staining Kit
Related Product Category Pages
- Fluorescent Mitcondria Stains
- Fluorescent Probes
- Enzyme Detection [Enzyme Substrates]
- Protein Stains Compatible with Western Blotting
- Blotting/Histochemistry Substrates
References
- 1) A Luminol/Iodophenol Chemiluminescent Detection System for Western Immunoblots
- 2) Quinone-based antibody labeling reagent for enzyme-free chemiluminescent immunoassays. Application to avidin and biotinylated anti-rabbit IgG labeling
- 3) Seeking Illumination: The Path to Chemiluminescent 1,2-Dioxetanes for Quantitative Measurements and In Vivo Imaging
