![]() ![]() Each containing different enhancers to increase signal, linearity and duration compared to other western blot detection systems. Compared to entry-substrates, SignalFire ECL Reagent boasts a more robust signal and extended duration of signal output. When I develop the blot with ECL select and image the. We offer six types of ECL substrates for western blot detection with horseradish peroxidase enzyme (HRP). SignalFire ECL Reagent from Cell Signaling Technology (CST) is an enhanced chemiluminescent substrate capable of detecting picogram amounts of protein by western blot analysis. I develop the membrane with a typical Western blot protocol with a HRP conjugated secondary antibody. Light emission occurs only during the enzyme-substrate reaction therefore, once the substrate in proximity to the enzyme is exhausted, signal output ceases. Total brain tissue was homogenized in RIPA lysis buffer containing protease and phosphatase inhibitors. It uses an enhanced chemiluminescent substrate for western blotting that was developed for film imaging and is also compatible with CCD imaging. When incubated with a blot on which HRP-conjugated antibodies (or other probes) are bound, a chemical reaction emits light at 425 nm which can be captured with x-ray film, CCD camera imaging devices and phosphorimagers that detect chemiluminescence. Product overview High Sensitivity ECL Substrate Kit ab133406 is designed for the detection of proteins with 23pg-187ng of protein per band. To make a working solution, the two components are mixed together. The two most common enzyme reporters that catalyze chemiluminescent reactions required for generating a recordable signal are horseradish peroxidase (HRP) and alkaline phosphatase (AP).ĮCL Substrates for High Sensitivity Western Blot DetectionĮnhanced chemiluminescence (ECL) substrates, are HRP substrates that are typically two-component systems consisting of a stable peroxide solution and an enhanced luminol solution. Recommended secondary antibody concentration: 1:50,000–1:250,000 dilution (4–20 ng/mL)Ĭhemiluminescent detection occurs when energy from a chemical reaction is released in the form of light.Compatibility: nitrocellulose and PVDF membranes.Stability: 24-hr working solution stability 1-year kit stability at room temperature. ![]() SuperSignal™ West Dura Features of Extended Duration Chemiluminescent Substrate ![]()
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