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SuperSignal West Femto Chemiluminescent Substrate


One of the most sensitive enhanced chemiluminescent substrates available.

SuperSignal West Femto Substrate for low femtogram detection levels

Thermo Scientific SuperSignal West Femto Maximum Sensitivity Substrate is an ultra-sensitive enhanced chemiluminescent (ECL) substrate for low-femtogram-level detection by Western blot analysis.

SuperSignal West Femto Substrate Substrate provides the ultimate sensitivity for Western blotting with horseradish peroxidase (HRP) enzyme. The substrate kit enables detection of low femtogram (that's mid-zeptomole!) amounts of protein on nitrocellulose or PVDF membrane when probed with appropriate primary and secondary antibodies. When combined with optimized antibody concentrations and blocking buffers, SuperSignal West Femto Substrate enables detection of target proteins in amounts that are too small to be seen with typical ECL substrates.

Highlights:

  • ECL – enhanced chemiluminescent substrate for horseradish peroxidase (HRP)
  • Sensitive – detect low-femtogram (mid-zeptomole) amounts of protein in bands on nitrocellulose or PVDF membranes
  • Quantitative – produces quantitative signal that is measurable over two orders of magnitude
  • Intense signal – easy to capture an image by exposure to film or imaging system
  • Excellent signal duration – 8 hours of usable light-output when conditions are optimized
  • Stable reagent – kit components are stable for 1 year at 4°C or 6 months at room temperature
  • Economical – optimized for extremely dilute antibody concentrations:
    • 1ng to 0.2µg/mL primary antibody (1:5000 to 1:100,000 dilution from 1mg/mL stock)
    • 2 to 10ng/mL secondary antibody (1:100,000 to 1:500,000 dilution from 1mg/mL stock)

Product Details:

SuperSignal West Femto Substrate provides femtogram-levels of protein detection in Western blots
True femtogram levels of detection with SuperSignal West Femto Maximum Sensitivity Substrate. Purified IκB was serially diluted from 100 to 1fg and then electrophoresed on a 4-20% mini gel. The protein was transferred to PVDF membrane and blocked withStartingBlock Blocking Buffer for 1 hour at room temperature. The blot was incubated in rabbit anti-IκBα (1mg/mL) at 1:1000 dilution overnight at 4°C, followed by incubation in HRP-conjugated goat anti-Rabbit IgG (1mg/mL) at 1:200,000 dilution for 1 hour at room temperature. The membrane was exposed to CL-XPosure Film for 1 minute.

SuperSignal West Femto Substrate is optimized for very dilute primary and secondary antibodies
Use less and see more with SuperSignal West Femto Substrate. A431cell lysate was diluted in electrophoresis reducing sample buffer at 5, 2.5, 1.25, 0.625, 0.3125, and 0.15625µg/well with a 10µL/well load. 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). The membrane was blocked with 5% milk in TBST and cut into three strips to optimize antibody dilutions. Membranes were then incubated with anti beta-Catenin (Part No. MA1-300) as indicated, followed by incubation with the appropriate dilution of Goat anti-Mouse Horseradish Peroxidase Conjugate (Part No. 31460). SuperSignal West Femto Substrate (Part No. 34095) was used for detection. Ten second exposures were acquired on CL-XPosure Film, 8 x 10 in. (Part No. 34091) and the myECL Imager (Part No. 62236). Exposures from the myECL Imager were inverted and contrasted (black=62,000, white=65,535, gamma=1.0).

Performance comparison between SuperSignal West Femto Substrate and other ECL substrates
Obtain maximum sensitivity and signal duration with SuperSignal West Femto Maximum Sensitivity Substrate. NIH3T3 lysate was diluted in electrophoresis reducing sample buffer. Lane 1 contained 5µg of NIH3T3 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-Erk1 (Part No. MA1-13041) at 0.2μg/mL and then with Goat anti-Mouse Horseradish Peroxidase Conjugate (Part No. 32430) at 5ng/mL. Identical blots were incubated in either SuperSignal West Femto Substrate (Part No. 34096), Clarity™ ECL Western Substrate (Bio-Rad Laboratories, Inc.), Luminata™ Forte Western HRP Substrate (EMD Millipore Corp.) or Immobilon™ Western Chemiluminescent HRP Substrate (EMD Millipore Corp.) according to each respective manufacturer's instructions. Two minute exposures of the resulting blots were simultaneously acquired on a single CL-XPosure Film, 8 x 10in. (Part No. 34091).

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
34094 SuperSignal West Femto 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 #34094 SuperSignal West Femto Substrate Trial Kit MSDS for product #34094 SuperSignal West Femto Substrate Trial Kit Certificate of Analysis for product #34094 SuperSignal West Femto Substrate Trial Kit $106.00

Add to cart
34095 SuperSignal West Femto Maximum Sensitivity 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 #34095 SuperSignal West Femto Maximum Sensitivity Substrate MSDS for product #34095 SuperSignal West Femto Maximum Sensitivity Substrate Certificate of Analysis for product #34095 SuperSignal West Femto Maximum Sensitivity Substrate $339.00

Add to cart
34096 SuperSignal West Femto Maximum Sensitivity 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 #34096 SuperSignal West Femto Maximum Sensitivity Substrate MSDS for product #34096 SuperSignal West Femto Maximum Sensitivity Substrate Certificate of Analysis for product #34096 SuperSignal West Femto Maximum Sensitivity Substrate $557.00

Add to cart
34095AB SuperSignal West Femto Maximum Sensitivity Substrate
Formulation: Proprietary luminol and peroxide solutions

Kit Contents:
Luminol/Enhancer
Stable Peroxide Buffer
Custom MSDS for product #34095AB SuperSignal West Femto Maximum Sensitivity Substrate

Instructions | MSDS | CofA
Product Instructions | MSDS | CofA  

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