Thermo Scientific™ Pierce™ ECL Western Blotting Substrate

The Thermo Scientific Pierce ECL Western Blotting Substrate is a reliable, cost-effective solution for detecting horseradish peroxidase (HRP) enzyme activity in Western blot applications using enhanced chemiluminescence (ECL). This substrate offers sensitivity, stability, and compatibility with standard membranes and imaging systems, providing robust performance for detecting picogram-level antigens.

Key Features

1. High Sensitivity:
   • Enables detection of low picogram levels of antigen, making it ideal for low-abundance protein analysis.
2. Ready-to-Use Formulation:
   • Two components: Luminol/Enhancer Solution and Stable Peroxide Buffer, pre-mixed at the time of use.
3. Long Signal Duration:
   • Chemiluminescent signal lasts 1 to 2 hours, providing ample time for imaging.
4. Stable and Long-Lasting:
   • Working solution remains stable for 1 hour.
   • Kit components are stable for 1 year at 4°C.
5. Broad Compatibility:
   • Compatible with nitrocellulose and PVDF membranes.
   • Compatible with commonly used imaging methods (e.g., photographic film or digital imaging systems).
6. No Re-Optimization Required:
   • Identical luminol and peroxide reagent formulations to other commercial ECL substrates, allowing seamless switching without changing protocols.
7. Economical Choice:
   • Provides equivalent performance to higher-cost ECL substrates (e.g., Amersham ECL Western Blotting Detection Reagent) at a value price.

Specifications

Available Quantities

Applications and Use Cases

1. Western Blotting for Protein Detection

• Purpose: Detect target proteins labeled with HRP-conjugated antibodies.
• Use Case: Researchers studying signaling pathways use Pierce ECL to detect phosphorylated proteins in cell lysates.

2. Low-Abundance Protein Detection

• Purpose: Enables detection of proteins present in low picogram quantities.
• Use Case: Detecting trace proteins in rare or precious biological samples, such as serum or tissue extracts.

3. High-Throughput Analysis

• Purpose: Supports large-scale protein detection on multiple blots.
• Use Case: Laboratories performing multiple Western blots for comparative studies or drug screening.

4. Reproducible Research

• Purpose: Provides consistent results across various experimental conditions.
• Use Case: Ensuring reproducibility in long-term research projects where standardized detection is critical.

5. Economical Routine Blotting

• Purpose: Offers a cost-effective alternative to expensive ECL substrates without compromising performance.
• Use Case: Routine blotting in teaching labs or for exploratory research.

Advantages

1. Ease of Use:
   • Ready-to-use components eliminate preparation errors and reduce handling time.
2. Cost-Effectiveness:
   • Provides performance comparable to premium substrates at a lower price.
3. Long Shelf Life:
   • Components remain stable for up to 1 year when stored correctly.
4. Broad Application:
   • Compatible with various membranes and detection systems, making it suitable for diverse experimental setups.
5. No Protocol Adjustments Needed:
   • Seamlessly replace costlier substrates without re-optimizing conditions.

Storage and Handling

• Store at 2–8°C to maintain stability.
• Product is shipped at ambient temperature and should be stored immediately upon receipt.
• Avoid repeated freeze-thaw cycles to ensure reagent integrity.

Recommended Protocol

1. Membrane Preparation:
   • Block the membrane after protein transfer with an appropriate blocking solution (e.g., 5% BSA or non-fat milk).
2. Antibody Incubation:
   • Incubate the membrane with the primary antibody (1:1,000 to 1:5,000 dilution) and then the HRP-conjugated secondary antibody (1:1,000 to 1:15,000 dilution).
3. Substrate Application:
   • Mix equal volumes of the Luminol/Enhancer and Stable Peroxide Buffer.
   • Apply sufficient substrate mixture to completely cover the membrane.
4. Imaging:
   • Capture the chemiluminescent signal using a photographic film or digital imaging system within 1–2 hours.

Why Choose Pierce ECL Western Blotting Substrate?

1. Proven Performance:
   • Delivers the same reliability as higher-cost alternatives (e.g., Amersham ECL) without the additional expense.
2. Enhanced Sensitivity:
   • Detects low picogram levels of protein for high-sensitivity applications.
3. Broad Compatibility:
   • Works with commonly used blotting membranes and imaging methods.
4. Long Signal Duration:
   • Provides ample time for signal acquisition and data capture.
5. Economical for Routine Use:
   • Ideal for laboratories performing high volumes of Western blotting.

The Thermo Scientific Pierce ECL Western Blotting Substrate offers robust, reliable, and cost-effective chemiluminescent detection, making it an excellent choice for both routine and sensitive Western blot applications. Let me know if you need additional details or assistance!