Enhanced chemiluminescence (ECL) reagents are used with horseradish peroxidase (HRP)-conjugated secondary antibodies to detect protein abundance in Western blot. HRP catalyzes luminol oxidation in the presence of hydrogen peroxide, producing light that can be detected by X-ray film and digital imagers.
PiQ™ ECL Substrate Kit is formulated with proprietary enhancers that greatly increase the intensity and duration of emitted light. PiQ™ ECL reagents are ultrasensitive and suitable to detect proteins at picogram level. When using these reagents, the recommended dilutions for primary and secondary antibodies are 1:5000 and 1:50,000, respectively. Compared to other commonly used ECL reagents, ours exhibit superior performance by generating extremely intense signal with low background and require ten-fold less primary and secondary antibodies.
Stable at 4°C for at least 1 year
- Allow the ECL reagents to equilibrate to room temperature approximately 30 min before use.
- Wash the membrane thoroughly to reduce non-specific signals.
- Mix Reagent A and B at a 1:1 ratio. Vortex to mix.
- Immerse the membrane in the mixed solution for 2–3 min.
- Drain the excess solution using a paper towel and wrap the membrane in plastic wrap or a clear plastic folder.
- Detect the chemiluminescence using X-ray film or a digital imager.