Chemiluminescence Western Blot - Reliable Chemiluminescent Protein Detection for Western Blots

Learn how to detect target proteins on western blots using chemiluminescent detection reagents such as SuperSignal™ Chemiluminescent HRP ECL Substrates: https://www.thermofisher.com/us/en/ho...

Watch More:
   • Western Blot Fundamentals  

What You'll Learn:
• How chemiluminescent detection works in western blotting
• When to use HRP-based ECL substrates
• How to apply detection reagents correctly
• Tips for maximizing signal sensitivity
• How to reduce background for clearer bands

Materials Used:
• SuperSignal™ Chemiluminescent HRP ECL Substrate
• Western blot membrane (PVDF or nitrocellulose)
• HRP-conjugated secondary antibody
• Wash buffer (e.g., TBST)
• Imaging system or film
• Incubation trays or containers

Steps Covered:
1. Wash the membrane after secondary antibody incubation.
2. Prepare the chemiluminescent substrate as directed.
3. Apply substrate evenly to the membrane.
4. Incubate briefly to allow signal development.
5. Detect protein signal using an imaging system or film.

CHAPTERS:
00:00 – Overview of chemiluminescent detection
00:30 – Prepare blot for detection
01:05 – Apply ECL substrate
01:45 – Signal development
02:20 – Image and visualize proteins

FAQs
Q: How do I choose the right ECL substrate sensitivity?
Select based on target abundance—higher sensitivity substrates are best for low-expression proteins.

Q: Can chemiluminescent substrates be reused?
No. Substrates should be prepared fresh for optimal signal performance.

Q: Is this compatible with digital imaging systems?
Yes, SuperSignal™ substrates are compatible with CCD and CMOS imagers.

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