Thermo Scientific SuperSignal West Dura Substrate is a luminol-based enhanced chemiluminescence (ECL) HRP substrate with very stable light output for femtogram-level detection with CCD-based imagers.
SuperSignal West Dura Extended Duration Substrate for HRP is optimized for high sensitivity and long signal duration, making it ideal for cooled-CCD camera detection systems. Unlike other substrates, whose signal declines to undetectable levels in 30 to 60 minutes, the signal produced with SuperSignal West Dura Substrate is stable for several hours, allowing multiple camera (or film) exposures to be made.
- ECL substrate – an enhanced chemiluminescent substrate for detection of horseradish peroxidase (HRP) activity from antibodies and other Western blot probes
- 24-hour signal – light emission is stable for 10 times longer than with typical ECL substrates; make multiple exposures to obtain publication-quality blot images
- Great sensitivity – provides femtogram-level detection; see bands you've never been able to see before
- Saves on antibody – works best with very dilute antibody concentrations, allowing you to perform 25 to 50 times more blots than with other chemiluminescent substrates
- High intensity – immediate stable signal generation provides easy detection with compatible imaging system (or x-ray autoradiography film)
- Reagent stability – 24-hour working solution stability; 1-year kit stability at room temperature
Thermo Scientific SuperSignal West Dura provides better sensitivity and less background than GE Healthcare ECL Plus* Substrate. Recombinant mouse IL-2 was serially diluted (50 to 0.003ng) and electrophoresed. The gel to be used for SuperSignal West Dura Substrate was transferred to nitrocellulose membrane and the gel to be used for ECL Plus Substrate was transferred to PVDF membrane (manufacturer's recommendation). The membranes were blocked and then incubated with a 1µg/mL dilution of the primary antibody, rat anti-mouse IL-2. After washing, the membranes were incubated with the secondary antibody, HRP-conjugated goat anti-rat IgG. The membranes were washed again and then incubated with substrates that were prepared according to the manufacturer's instructions. Each membrane was exposed to X-ray film for 5 minutes. The SuperSignal West Dura Substrate membrane was exposed to a ChemiImager* 4000 (Alpha Innotech) for 30 minutes and the ECL Plus blot was exposed for 15 minutes. (The exposure of the ECL Plus blot was not extended to 30 minutes due to the high background that had already accumulated at 15 minutes.)
||24-hour signal duration and high intensity. A dot blot with replicates of 37pg of horseradish peroxidase was incubated for 5 minutes in SuperSignal West Dura Substrate. Images were obtained using a ChemiImager* 4000 (Alpha Innotech) at various times after removing the blot from the substrate. The image exposure period is indicated for each time point.
Along with the advances in cooled CCD technology, lower cost of imaging systems and the desire to eliminate chemical waste associated with film development, it is increasingly common to see imaging instruments in research labs. Cooled CCD cameras allow instant image manipulation, higher sensitivity and a larger dynamic range than film. Use of imagers eliminates the need for film processing equipment and darkrooms. However, such systems require substrates that produce intense, long-duration signal (light-emission) to be captured effectively by the camera sensor. SuperSignal West Dura Extended Duration Substrate was developed to meet this need, combining 24-hour light emission with very high intensity, allowing researchers to take full advantage of all the features offered by imaging instruments.
Learn more about chemiluminescent Western blot detection
Watch the Western blot video tutorials
Pierce Fast Western Blot Kits with SuperSignal Substrates
Selection Guide for Chemiluminescent Western Blot Substrates
Blotting Equipment, Membranes and Signal Enhancers
CL-XPosure Film (X-ray film for chemiluminescence)