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Western Blot (Sigma Protocol) moderate

Inactive Chemiluminescent Detection

Symptom
No signal detected when using chemiluminescent detection method despite proper procedure
Common Causes
  1. 1 Film development solution has expired
  2. 2 Chemiluminescent substrate prepared incorrectly
  3. 3 Fresh H2O2 not added to substrate when required
Solutions
  1. 1 Use fresh film development solutions
  2. 2 Follow substrate manufacturer's instructions carefully
  3. 3 Ensure fresh H2O2 is added if required by substrate protocol
Related Video (3)
Cell Signaling Technology ★ 85
Western Blot Troubleshooting Guide
"Directly addresses troubleshooting Western blot signal failure, including detection issues."
Bilibili (China-Accessible Mirrors) ★ 75
Western blot full protocol: Protein extraction to chemiluminescence
"Shows the complete Western blot workflow including chemiluminescence detection step where signal would appear."
Bilibili (China-Accessible Mirrors) ★ 70
Western Blot Complete Process First-Person View
"Provides a first-person complete protocol to verify all steps, useful for checking reagents like film developer."
Source: sigmaaldrich.com ↗
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