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SuperSignal Western Blot Enhancer


Increase signal-to-noise ratio and band development for better sensitivity.

SuperSignal Western Blot Enhancer increases Western blot sensitivity and reduces background.

The Thermo Scientific SuperSignal Western Blot Enhancer contains a membrane treatment reagent and a primary antibody diluent that increase both signal intensity and sensitivity 3- to 10-fold compared to detection performed without it.

When a protein or antigen is difficult to detect because of low abundance or poor immunoreactivity, use of SuperSignal Western Blot Enhancer can significantly reduce background and enhance detection of low-abundance and weakly immunoreactive antigens. This versatile kit works with PVDF and nitrocellulose membranes and is compatible with fluorescent, chromogenic and chemiluminescent detection.

Highlights:

  • Increase sensitivity – achieve 3- to 10-fold increase in signal intensity and sensitivity
  • Improve specificity – improves signal-to-noise ratio for poor quality and low affinity antibodies
  • Better clarity – reduces background for cleaner Western blots
  • Membrane compatibility – provides effective signal enhancement with PVDF and nitrocellulose membranes
  • Substrate compatibility – validated for use with chromogenic, chemiluminescent and fluorescent detection methods

Product Details:


SuperSignal Western Blot Enhancer reduces background to produce cleaner blots and allow longer exposure times.
 
Thermo Scientific SuperSignal Western Blot Enhancer reduces background to enable detection of low-abundance targets with low abundance. K562 cell lysate was loaded into Tris-glycine SDS-PAGE gels at 1.25, 2.5, 5 and 10µg per lane. (Left) After electrophoresis the proteins were transferred to nitrocellulose membranes (Part No. 88013) and the membrane blocked with Thermo Scientific SuperBlock Blocking Buffer (Part No. 37535). The blot was probed with mouse anti-ERK 1 (Part No. MA1-13041) primary antibody at 1µg/mL and then a horseradish peroxidase-conjugated goat anti-mouse IgG (Part. No. 31430) secondary antibody at 0.08µg/mL. Thermo Scientific Pierce ECL Substrate (Part No. 32209) was used for detection. Both the primary antibody and secondary antibody conjugate were diluted in blocking buffer. (Right) As described above except using the SuperSignal Western Blot Enhancer protocol. Membranes were pretreated with the Antigen Pretreatment Solution for 10 minutes, rinsed with water and incubated with blocking buffer. The primary antibody was diluted in Primary Antibody Diluent prior to probing the membrane.

 


SuperSignal Western Blot Enhancer improves the lower detection limit of antibodies and increases the sensitivity of Western Blots for low-abundance targets.
 
Thermo Scientific SuperSignal Western Blot Enhancer improves the lower detection limit for chemiluminescent substrates. Cell lysates were separated by electrophoresis, transferred to nitrocellulose and Western blotting was performed using the conventional method (Left) or using the SuperSignal Western Blotting Enhancer protocol (Right). Membranes with PC-3 lysate (Top) were blocked with 5% milk in Tris-Buffered Saline with 0.05% Tween™ 20 and probed with mouse anti-CDK 5 (LabVision Part No. MS-1059-PABX) at 1µg/mL followed by horseradish peroxidase conjugated goat anti-mouse IgG (Part No. 31430) at 0.1µg/mL. Detection was performed with Pierce ECL Substrate (Part No. 32209) and exposed to film for 1 minute. Membranes with HeLa lysate (Bottom) were blocked with Thermo Scientific Blocker BLOTTO in TBS (Part No. 37530) and probed with rabbit anti-GSK3α (Cell Signaling Technology Part No. 9338) at 1µg/mL followed by horseradish peroxidase-conjugated goat anti-rabbit IgG (Part No. 31460) at 0.1µg/mL. Detection was performed with SuperSignal West Pico Chemiluminescent Substrate (Part No. 34080) and exposed to film for 10 minutes.

 


SuperSignal Western Blot Enhancer for Chromogenic and Fluorescent Western Blots
 
Thermo Scientific SuperSignal Western Blot Enhancer is compatible with chromogenic and fluorescent detection methods. HeLa cell lysate was separated by electrophoresis, transferred to nitrocellulose (Top) or Low-Fluorescence PVDF (Part No. 22860) (Bottom) and Western blotting was performed using the conventional method (Left) or using the SuperSignal Western Blot Enhancer protocol (Right). For chromogenic detection (Top), the nitrocellulose membranes were blocked with SuperBlock in TBS (Part No. 37535) and probed with rabbit anti-β-Catenin (LabVision Part No. PAb RB-1491-PABX) at 0.2µg/mL followed by alkaline phosphatase-conjugated goat anti-rabbit IgG (Part No. 31340) at 0.04µg/mL. Detection was performed with Thermo Scientific 1-Step NBT/BCIP Substrate (Part No. 34042). For fluorescent detection (Bottom) , the PVDF membranes were blocked with Blocker BLOTTO in TBS (Part No. 37530) and probed with rabbit anti-MAP Kinase at 1µg/mL followed by Thermo Scientific DyLight 488-conjugated and goat anti-rabbit IgG (Part No. 35552) at 0.1µg/mL. Membranes were imaged with the GE Healthcare Typhoon™ 9410 Instrument.

 


SuperSignal Western Blot Enhancer compared to SignalBoost and Chemilucent Plus Products
 
Comparison of Thermo Scientific SuperSignal Western Blot Enhancer with other commercially available enhancers. HeLa cell lysate was separated by electrophoresis, transferred to nitrocellulose (Part No. 88013) and Western blotting was performed using the conventional method (A), SuperSignal Western Blot Enhancer (B), EMD Biosciences SignalBoost™ Immunoreaction Enhancer Kit (C) or Millipore Chemilucent™ Plus Western Blot Enhancing Kit (D) according to manufacturers' directions. Blots were probed with rabbit anti-MAP Kinase (Millipore Part No. 06-182) at 1µg/mL followed by horseradish peroxidase-conjugated goat anti-rabbit IgG (Part No. 31460) at 0.1µg/mL. Detection was performed with Pierce ECL Substrate (Part No. 32209) and exposed to film for 10 minutes.

Related Products:

Blocking Buffers
Western Blotting Substrates
Blotting Equipment, Film and Membranes


Ordering Information
 
Product # Description Pkg. Size Instructions MSDS CofA Price
46640 SuperSignal Western Blot Enhancer
Formulation: Proprietary clear, colorless solutions
Sufficient For: 25 mini blots or 2000cm2 membrane


Kit Contents:
Antigen Pretreatment Solution, 250mL
Primary Antibody Diluent, 250mL
500mL kit Product Instructions for product #46640 SuperSignal Western Blot Enhancer MSDS for product #46640 SuperSignal Western Blot Enhancer Certificate of Analysis for product #46640 SuperSignal Western Blot Enhancer $213.00

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46641 SuperSignal Western Blot Enhancer
Formulation: Proprietary clear, colorless solutions
Sufficient For: 2 mini blots or 160cm2 membrane


Kit Contents:
Antigen Pretreatment Solution, 25mL
Primary Antibody Diluent, 25mL
50mL kit Product Instructions for product #46641 SuperSignal Western Blot Enhancer MSDS for product #46641 SuperSignal Western Blot Enhancer Certificate of Analysis for product #46641 SuperSignal Western Blot Enhancer $44.00

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Instructions | MSDS | CofA
Product Instructions | MSDS | CofA  

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