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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)
Bilibili (China-Accessible Mirrors) ★ 88
Western blot full protocol: Protein extraction to chemiluminescence
"Full workflow explicitly includes chemiluminescence detection, directly showing the step where expired developer causes no signal."
Cell Signaling Technology ★ 80
Western Blot Troubleshooting Guide
"Troubleshooting guide specifically covers diagnosing no-signal Western blot problems including detection reagent issues."
Bilibili (China-Accessible Mirrors) ★ 72
Western blot hands-on: Protein extraction through detection
"Hands-on protocol with troubleshooting emphasis covers pitfalls across extraction through detection including chemiluminescent development."
Source: sigmaaldrich.com ↗
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