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SuperSignal West Dura Chemiluminescent Substrate


Long signal duration and high signal intensity, ideal for CCD cameras.

SuperSignal West Dura Chemiluminescent Western Blot Substrate for long-duration, stable light emission.

Thermo Scientific SuperSignal West Dura Extended Duration Substrate is a luminol-based enhanced chemiluminescence (ECL) HRP substrate with stable light output for mid-femtogram-level detection with CCD-based imagers.

SuperSignal West Dura 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.

Highlights:

  • ECL – enhanced chemiluminescent substrate for horseradish peroxidase (HRP)
  • 24-hour signal duration – light emission is stable for 10 times longer than with typical ECL substrates; acquire multiple exposures to obtain publication-quality blot images
  • Great sensitivity – provides detection of mid-femtogram levels of target proteins
  • High intensity – immediate, stable signal generation provides easy detection via film or imager
  • Stable reagent – 24-hour working solution stability; 1-year kit stability at room temperature
  • Saves on antibody – works best with dilute antibody concentrations:
    • 20ng to 1.0µg/mL primary antibody (1:1000 to 1:50,000 dilution from 1mg/mL stock)
    • 4 to 20ng/mL secondary antibody (1:50,000 to 1:250,000 dilution from 1mg/mL stock)

Product Details:

SuperSignal West Dura Substrate compared to Bio-Rad Clarity Western ECL Substrate
Obtain mid-femtogram detection with SuperSignal West Dura Substrate. Turbo GFP-His-HA-FLAG was diluted in electrophoresis reducing sample buffer. Lane 1 contained 1,000 fg of Turbo GFP. Seven 1:1 serial dilutions were then prepared and applied at 10μL/well. After electrophoresis, proteins were transferred to PVDF membrane (Part No. 88518), and then the membrane was blocked with SuperBlock in TBST Buffer (Part No. 37536). The membrane was incubated with Biotinylated Anti-His (Part No. MA1-21315-BTIN) at 1μg/mL and then with Streptavidin-Horseradish Peroxidase Conjugate (Part No. 21126) at 1:50,000 dilution. SuperSignal West Dura Substrate (Part No. 34076) was used for detection.

SuperSignal West Dura Substrate compared to Amersham ECL Prime
Longer signal duration with SuperSignal West Dura 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 Amersham™ 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 Substrate compared to Amersham ECL Prime
Better sensitivity and signal duration with SuperSignal West Dura Substrate. A431 cell lysate was diluted in electrophoresis reducing sample buffer. Lane 1 contained 5µg of A431 lysate. Five 1:1 serial dilutions were then prepared and applied at 10μL/well. After electrophoresis, proteins were transferred to nitrocellulose membrane (Part No. 88018) using the Pierce G2 Fast Blotter (Part No. 62288) and Pierce 1-Step Transfer Buffer (Part No. 84731). Membranes were blocked with 5% milk in TBST Buffer. The membranes were incubated with Anti-beta-catenin (Part No. MA1-300) at 0.3μg/mL and then with Goat anti-Mouse Horseradish Peroxidase Conjugate (Part No. 32430) at 20ng/mL. Identical blots were incubated in either SuperSignal West Dura Substrate (Part No. 34076), Clarity™ ECL Western Substrate (Bio-Rad Laboratories, Inc.) or Immobilon™ Western Chemiluminescent HRP Substrate (EMD Millipore Corp.) according to respective manufacturer's instructions. Thirty second exposures of the resulting blots were simultaneously acquired on the myECL Imager (Part No. 62236) with the following settings: black = 65,296, white = 65,535, and gamma = 1.0).

SuperSignal West Dura Substrate compared to Amersham ECL Prime
Product comparison. Blots detected using Thermo Scientific SuperSignal West Dura Substrate (top) or Luminata™ Crescendo Western HRP Substrate (EMD Millipore Corp.) (bottom) at 10 minutes post substrate incubation. Bands are beta-catenin in HeLa cell lysate samples (10µg total protein in first lane; 2-fold dilutions thereafter), probed with rabbit anti-beta-catenin antibody (Part No. RB-1491-PABX) at 1:2000 dilution, followed by goat anti-rabbit HRP antibody (Part No. 31460) at 4ng/mL and exposure to CL-XPosure X-ray film (Part No. 34091).

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).

More Product Data

Related Resources:

Western blotting and detection technical guide
Western blot video tutorials

Related Products:

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)
myECL Imager


Ordering Information
 
Product # Description Pkg. Size Instructions MSDS CofA Price
37071 SuperSignal West Dura Substrate Trial Kit
Formulation: Proprietary luminol and peroxide solutions
Sufficient For: 2 mini-gel blots; 200cm2 of membrane

Kit Contents:
Luminol/Enhancer, 10mL
Stable Peroxide Buffer, 10mL
20mL kit Product Instructions for product #37071 SuperSignal West Dura Substrate Trial Kit MSDS for product #37071 SuperSignal West Dura Substrate Trial Kit Certificate of Analysis for product #37071 SuperSignal West Dura Substrate Trial Kit $100.00

Add to cart
34075 SuperSignal West Dura Extended Duration Substrate
Formulation: Proprietary luminol and peroxide solutions
Sufficient For: 12 mini-gel blots; 1000cm2 of membrane

Kit Contents:
Luminol/Enhancer, 50mL
Stable Peroxide Buffer, 50mL
100mL kit Product Instructions for product #34075 SuperSignal West Dura Extended Duration Substrate MSDS for product #34075 SuperSignal West Dura Extended Duration Substrate Certificate of Analysis for product #34075 SuperSignal West Dura Extended Duration Substrate $320.00

Add to cart
34076 SuperSignal West Dura Extended Duration Substrate
Formulation: Proprietary luminol and peroxide solutions
Sufficient For: 25 mini-gel blots; 2000cm2 of membrane

Kit Contents:
Luminol/Enhancer, 100mL
Stable Peroxide Buffer, 100mL
200mL kit Product Instructions for product #34076 SuperSignal West Dura Extended Duration Substrate MSDS for product #34076 SuperSignal West Dura Extended Duration Substrate Certificate of Analysis for product #34076 SuperSignal West Dura Extended Duration Substrate $535.00

Add to cart
34075AB SuperSignal West Dura Extended Duration Substrate
Formulation: Proprietary luminol and peroxide solutions

Kit Contents:
Luminol/Enhancer
Stable Peroxide Buffer
Custom MSDS for product #34075AB SuperSignal West Dura Extended Duration Substrate

Instructions | MSDS | CofA
Product Instructions | MSDS | CofA  

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