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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 Substrate is an ultra-sensitive enhanced chemiluminescent (ECL) substrate for low-femtogram-level detection by Western blot analysis.

SuperSignal West Femto Substrate Maximum Sensitivity Substrate is an ultra-sensitive chemiluminescent substrate system for Western blotting with horseradish peroxidase (HRP) enzyme. The substrate kit enables detection of low femtogram (that's mid-zeptomole!) amounts of protein deposited on nitrocellulose or PVDF membrane and 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 small to be seen with typical ECL substrates.

Highlights:

  • ECL substrate – an enhanced chemiluminescent substrate for detection of horseradish peroxidase (HRP) activity from antibodies and other Western blot probes
  • 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
  • Saves on antibody – optimized for very dilute antibody concentrations; dilute primary antibodies 5000- to 100,000-fold and secondary antibodies 100,000 to 500,000-fold (from 1mg/mL stocks)
  • Intense signal – easy to capture an image by exposure to autoradiography film or via a CCD camera or similar 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

Product Details:


SuperSignal West Femto Substrate provides true femtogram-levels detection
 
True femtogram levels of detection with Thermo Scientific 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 with Thermo Scientific StartingBlock 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 Thermo Scientific CL-XPosure Film for 1 minute.


SuperSignal West Femto Substrate is optimized for very dilute primary and secondary antibodies
 
Use less antibody with Thermo Scientific SuperSignal West Femto Maximum Sensitivity Substrate. MAP Kinase and HSP86 were detected in HeLa cell lysate prepared by dilution in Laemmli sample buffer and heating to 95°C for 5 minutes. Serial dilutions (1:1) of total protein (10µg to 19.5ng) were separated by electrophoresis and the proteins transferred to nitrocellulose membranes (Part No. 88013). Thermo Scientific Starting Block in TBS Blocking Buffer (Part No. 37542) was used for blocking. Rabbit anti-MAP Kinase (Millipore Part No. 06-182) and Rabbit anti-HSP86 (Part No. PA3-013) were used as primary antibodies at 0.2µg/mL (1:5000 dilution of 1mg/mL stock solution). Goat anti-Rabbit HRP (Part No. 31460) was used as secondary antibody at 2ng/mL (1:500,000 dilution of 1mg/mL stock solution). MAP Kinase and HSP86 targets were detected using Thermo Scientific SuperSignal West Femto (Part No. 34095) and Thermo Scientific CL-XPosure Film (Part No. 34091).


SuperSignal West Femto Substrate is more sensitive than ECL Plus Substrate
 
Thermo Scientific SuperSignal West Femto is more sensitive than GE Healthcare ECL Plus Substrate. Two-fold serial dilutions of mouse IL-2 were detected with both substrates. The primary antibody dilution was 1/2000 and the secondary antibody dilution was 1/300,000 (from 1mg/mL stocks).

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)


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
34095PROMO SuperSignal West Femto Maximum Sensitivity Substrate
Formulation: 4 x Part No. 34095

Promotional item; expires June 30, 2014
4 x 100mL kit $1,017.00

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
34096PROMO SuperSignal West Femto Maximum Sensitivity Substrate
Formulation: 4 x Part No. 34096

Promotional item; expires June 30, 2014
4 x 200mL kit $1,671.00

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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