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 similar sensitivity and more stable signal duration compared to Amersham™ ECL Prime Substrate. Detection of Hsp86 in HeLa cell lysate on nitrocellulose membranes. HeLa cell lysate was diluted in sample buffer and heated to 95˚C for 5 minutes. Lane 1 contained 10μg total protein (total 10μL/well). Four 1:1 serial dilutions were then prepared and loaded at 10μL/well (Lanes 2-5). After transfer to nitrocellulose membranes (Part No. 88024), blots were blocked (Part No. 37542), probed with rabbit anti-Hsp 86 antibody (Part No. PA3-013) at 1:2000 dilution, followed by goat anti-rabbit IgG HRP secondary antibody (Part No. 31460) at 6.6ng/mL. Finally, respective blots were incubated with SuperSignal West Dura Substrate (Part No. 34076) or ECL Prime (GE Healthcare, Part No. RPN2232) per product instructions. At various time-points following substrate incubation, the two blots were imaged using a CCD camera imager. (Identical imaging exposure parameters were used for both blots at each time-point.)
|SuperSignal West Dura is more sensitive than Bio-Rad™ Clarity™ Western ECL Substrate. Detection of MAP Kinase (Erk 1/2) and Hsp86 proteins in HeLa cell lysate on PVDF membranes. Membranes were probed, processed and exposed to X-ray film using identical conditions, except that incubations in substrates were according to respective manufacturer's product instructions.
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. SuperSignal West Dura Extended Duration Substrate was developed for use with cooled CCD imaging systems, such as the Thermo Scientific myECL Imager (Part No. 62236).
Western blotting and detection technical guide
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)