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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
"Direct troubleshooting guide for Western blotting that will address detection failures and reagent-related issues including chemiluminescent detection problems."
Bilibili (China-Accessible Mirrors) ★ 85
Western blot full protocol: Protein extraction to chemiluminescence
"Complete workflow explicitly covering chemiluminescence detection step, allowing researcher to verify proper procedure and identify where expired reagents impact signal generation."
Cell Signaling Technology ★ 78
Western Blotting Protocol
"Comprehensive Western blot protocol emphasizing reproducible and reliable results, including detection methodology that would expose gaps when reagents are expired."
Source: sigmaaldrich.com ↗
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