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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 ★ 92
Western Blot Troubleshooting Guide
"Dedicated Western blot troubleshooting guide that directly addresses no-signal detection failures and their causes."
Bilibili (China-Accessible Mirrors) ★ 78
Western blot full protocol: Protein extraction to chemiluminescence
"Full protocol including chemiluminescence detection, showing the reagents and step where expired substrate causes no signal."
Bilibili (China-Accessible Mirrors) ★ 72
Western blot hands-on: Protein extraction through detection
"Step-by-step protocol with emphasis on troubleshooting common pitfalls including detection issues."
Source: sigmaaldrich.com ↗
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