Background

Enhanced chemiluminescence (ECL) substrates are used with horseradish peroxidase (HRP)-conjugated secondary antibodies to detect protein abundance in immunoassays. HRP catalyzes luminol oxidation in the presence of hydrogen peroxide, producing light that can be detected by an X-ray film, digital imager or chemiluminescence plate reader.

PiQ™ ECL Substrate Kit is formulated with proprietary enhancers that significantly increase the intensity and duration of light emitted. The PiQ™ ECL reagents are ultrasensitive and suitable for detecting proteins at the picogram level. When using these reagents in Western blotting, the recommended dilutions for primary and secondary antibodies are 1:5,000 and 1:50,000, respectively. We also tested this product in chemiluminescent ELISA, it outperformed other commercially available ECL reagents by generating much stronger signals with a relative low background.

Storage

Stable at 4°C for at least 1 year

Experimental Protocol

Let the ECL reagents warm up at room temperature for approximately 30 min prior to use.

Combine Reagents A and B at a ratio of 1:1. Vortex to mix.

Apply the mixture to the Western blots or microplate wells.Immediately capture signals with a digital imager or chemiluminescence plate reader.

Application Tested

Western blotting

Chemiluminescent ELISA

Product Features

Superior performance in generating extremely intense signals

Suitable for detecting low-abundance analytesOptimized for Western blotting and chemiluminescent ELISA