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  5. Thermo Scientific™ Pierce™ ECL Western Blotting Substrate

Thermo Scientific™ Pierce™ ECL Western Blotting Substrate

Catalog No. FER32209X4
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Description

Value-priced, entry-level peroxidase substrate for enhanced chemiluminescence (ECL) that directly replaces costlier products without the need to re-optimize conditions.

Thermo Scientific Pierce ECL Western Blotting Substrate is a value-priced, entry-level horseradish peroxidase (HRP) substrate for enhanced chemiluminescence (ECL) that directly replaces costlier products without the need to re-optimize conditions.

Pierce ECL Plus Western Blotting Substrate characteristics include:

  • Sensitivity: low picogram
  • Stability: 1-hr working solution stability, 1-year kit stability at 4°C
  • Compatibility: nitrocellulose and PVDF membranes
  • Signal duration: 1 to 2 hours
  • Recommended primary antibody concentration: 1:1,000–1:5,000 dilution (0.2–10 μg/mL)
  • Recommended secondary antibody concentration: 1:1,000–1:15,000 dilution (0.07–1.0 μg/mL)

Pierce ECL Western Blotting Substrate provides reliability and performance equivalent to other standard ECL substrates for detection of HRP enzyme activity. It enables the detection of picogram amounts of antigen and allows for easy detection of HRP using photographic or other imaging methods.

Because the luminol and peroxide reagent formulations are identical to other commercially available substrate products, one can switch to Pierce ECL without needing to optimize probing conditions or incubation protocols. Pierce ECL Western Blotting Substrate provides performance identical to the original Amersham ECL Western Blotting Detection Reagent from GE Healthcare.

Specifications

Product Type Substrate
Detection Method Chemiluminescence
Form Liquid
Product Line Pierce
Quantity 4 x 250 mL
Substrate Horseradish Peroxidase
Recommended Antibody Concentrations 2°: 1:1-15 K (0.07-1 μg/mL), 1°: 1:1 K (0.2-10 μg/mL)
Sensitivity Low picogram
Signal Duration 1 to 2 hr.
Kit Contents Luminol/Enhancer, 4 x 125 mL, Stable Peroxide Buffer, 4 x 125 mL
Content And Storage Upon receipt store at 2-8°C. Product is shipped at ambient temperature.
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Frequently Asked Questions (FAQs)

How can I increase the sensitivity of my Western blot?

Consider transferring to a different membrane or using a different detection method. We have observed increased sensitivity when using PVDF membranes in place of nitrocellulose. On PVDF membranes, using as little as 1 µg of total rat brain protein, PKC can be detected with alkaline phosphatase-mediated chromogenic detection in some cases using affinity-purified antibodies at a concentration of 0.5 µg/mL. Detection sensitivity can also be increased by using chemiluminescent detection, especially when using a SuperSignal West enhanced chemiluminescence subtrate (https://www.thermofisher.com/us/en/home/life-science/protein-biology/protein-assays-analysis/western-blotting/detect-proteins-western-blot/western-blot-detection-reagents/detection-technologies-western-blotting/chemiluminescent-western-blot-detection/supersignal-chemiluminescent-substrates.html) such as SuperSignal West Pico PLUS, SuperSignal West Dura, or SuperSignal West Femto. The secondary antibody should be used as recommend by the manufacturer and optimized as needed.

For Research Use Only. Not for use in diagnostic procedures.