ECL Substrates for Western Blotting
Clarity and Clarity Max are compatible with any horseradish peroxidase (HRP) conjugate and ideal for all western blots with both digital and film-based imaging systems.View ECL Blotting Substrates
ECL Substrates for High-Sensitivity Western Blot Detection
Chemiluminescent assays depend on the emission of light as a product of a chemical reaction and are commonly used for the detection of proteins on western blots. The secondary antibody used in western blotting is conjugated to the horseradish peroxidase (HRP) enzyme which reacts with the HRP substrate luminol. This reaction emits light at 428 nm and this light signal can be captured on X-ray film or by a digital imager.
However, luminol emits light only weakly, so enhancers are added to the reaction to increase the signal. This enhancement of a luminol-based signal is commonly referred to as enhanced chemiluminescence (ECL).
There are a number of different enhancers available, some of which can increase the signal by as much as a 1,000-fold, making ECL more sensitive than other common detection systems such as conversion of substrates to colored precipitates. For example, the enhancers used in Bio-Rad Clarity Max ECL substrate maximizes detection sensitivity to enable detection to the low femtogram amounts of target. The type of enhancer used can also increase the signal duration. Luminol has an emission half-life of less than a minute. The Bio-Rad Clarity ECL substrate extends the signal duration to 24 hours, permitting reimaging without loss of signal intensity.
- Sensitive detection down to the femtogram level of target
- Low background signal for cleaner looking blots
- Long signal duration for re-imaging flexibility
- Compatibility with digital or film-based imaging
- Compatibility with any HRP conjugate
- Long shelf life at room temperature
Western blotting products include the V3 Western Workflow, protein transfer systems, membranes, filter paper, blotting buffers, and detection kits
Choose SDS-PAGE and native PAGE gels, convert to TGX precast gels, or choose specialized gel chemistries including Stain-Free
Detect, image and quantify multiplex fluorescence, chemiluminescence, stain-free and colorimetric blots and gels.
Immunodetection (immunological detection) is used to identify specific proteins blotted to membranes. This section provides an overview of immunodetection methods, workflow, protocol, and troubleshooting tips.