Home › Failure Case Library › Inactive Chemiluminescent Detection
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)
Bilibili (China-Accessible Mirrors) ★ 92
Western blot full protocol: Protein extraction to chemiluminescence
"Explicitly covers chemiluminescence detection step where expired reagents would cause signal failure; shows complete workflow including the critical detection phase"
Cell Signaling Technology ★ 88
Western Blot Troubleshooting Guide
"Dedicated troubleshooting guide for Western blotting that directly addresses no-signal problems and their root causes"
Bilibili (China-Accessible Mirrors) ★ 85
Western Blot — Full Protocol Walkthrough (Bio-Techne)
"Comprehensive protocol demonstration including ECL detection phase where expired development solution problems would manifest"
Source: sigmaaldrich.com ↗
← Back to all cases