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Thermo Scientific™ Pierce™ ECL Plus Western Blotting Substrate

Catalog No. PI32134
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25 mL Kit
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PI32134 25 mL Kit
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Catalog No. PI32134 Supplier Thermo Scientific™ Supplier No. 32134
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Acridan-based chemiluminescent and chemifluorescent HRP substrate for Western blot detection using X-ray film or CCD- or laser-based imagers.

Thermo Scientific™ Pierce ECL Plus Substrate is an acridan-based chemiluminescent and chemifluorescent substrate to detect peroxidase activity from HRP-conjugated probes by Western blotting using X-ray film, CCD or laser-based imagers.

Thermo Scientific Pierce ECL Plus Substrate is an acridan-based chemiluminescent and chemifluorescent HRP substrate for western blot detection using X-ray film or CCD- or laser-based imagers. This product is sold as Pierce ECL 2 Substrate (Cat. #PI80196X3) through Fisher Scientific and other channels.

Features of ECL Plus Substrate:

  • Easy to use—can be substituted for the discontinued GE ECL Plus Substrate without any re-optimization
  • Higher sensitivity—detect targets down to the low-picogram level
  • Longer signal duration—sustained light output for as long as 5 hours
  • More imaging options—X-ray, CCD or laser-based imagers
  • More affordable—high quality and performance at a lower price than other suppliers' similar substrates

Pierce ECL Plus Substrate generates acridinium esters when it reacts with HRP. As these ester intermediates react with peroxide, they produce a strong and sustained chemiluminescent signal that can be captured by X-ray film CCD imaging. The substrate also produces a robust fluorescent signal that can be captured with fluorescence imagers. Pierce ECL Plus Substrate enables the detection of low-picogram amounts of target protein on nitrocellulose or PVDF membrane when probed with appropriate primary and secondary antibody concentrations. Its broad dynamic range, high sensitivity and long-lasting signal make Pierce ECL Plus Substrate an excellent choice for western blot analysis.

Specifications

Product Type Substrate
Detection Method Chemiluminescence, Fluorescence
Form Liquid
For Use With (Application) Western Blot
Membrane Compatibility Nitrocellulose, PVDF
Product Line Pierce
Quantity 25 mL Kit
Substrate Horseradish Peroxidase
Recommended Antibody Concentrations 2°: 1:25-200K (5-40ng/mL), 1°: 1:1K (0.05- 1μg/mL)
Sensitivity Low picogram to mid-femtogram
Signal Duration 5 hr.
Sufficient For 250 cm2 of membrane
Kit Contents Detection Reagent A, 25 mL, Detection Reagent B, 0.625 mL
Content And Storage Upon receipt store at 4°C. Product shipped on ice.
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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.