Chemiluminescent detection occurs when energy from a chemical reaction is released in the form of light. The two most common enzyme reporters that catalyze chemiluminescent reactions required for generating a recordable signal are horseradish peroxidase (HRP) and alkaline phosphatase (AP).
Enhanced chemiluminescence (ECL) substrates, are HRP substrates that are typically two-component systems consisting of a stable peroxide solution and an enhanced luminol solution. To make a working solution, the two components are mixed together. 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. Light emission occurs only during the enzyme-substrate reaction; therefore, once the substrate in proximity to the enzyme is exhausted, signal output ceases.
We offer six types of ECL substrates for western blot detection with horseradish peroxidase enzyme (HRP). Each containing different enhancers to increase signal, linearity and duration compared to other western blot detection systems.
SuperSignal West Atto | SuperSignal West Pico Plus | SuperSignal West Dura | |
Choose when? | Target is very low-abundant, sample is limited, or antibodies are limited | Everyday applications; offers improved sensitivity over base level ECL | Performing quantitative western blotting or maximum signal duration is needed |
Features | High Sensitivity | Widest dynamic range | Longest duration and linearity |
Sensitivity | Low femtogram to high attogram | Low picogram to femtogram | Mid femtogram |
Duration | 6 hours | Up to 24 hours | 24 hours |
Recommend antibody dilutions (1 mg/ml stock) |
1°: 1:5,000 2°: 1:100,000-1:250,000 |
1°: 1:1,000 2°: 1:20,000-1:100,000 |
1°: 1:5,000 2°: 1:50,000-1:250,000 |
Catalog No | A38554 (20 mL) A38555 (100 mL) A38556 (200 mL) |
34579 (20 mL) 34577 (200 mL) 34580 (500 mL) 34578 (1 L) |
37071 (20 mL) 34075 (100 mL) 34076 (200 mL) |