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Immunohistochemistry (No Staining) severe

Antibody not validated for IHC application or tissue type

Symptom
Complete absence of staining with antibody that works in other applications. The antibody may not recognize native protein conformation present in tissue sections.
Common Causes
  1. 1 Antibody designed for denatured proteins (western blot) not recognizing native epitopes in IHC
  2. 2 Antibody not validated for specific tissue preparation method (FFPE vs. fresh frozen)
  3. 3 Epitope only accessible in denatured state, not in tissue architecture
  4. 4 Antibody specificity incompatible with IHC application requirements
Solutions
  1. 1 Check antibody datasheet to confirm IHC validation
  2. 2 Verify antibody is validated for specific tissue preparation (FFPE, fresh frozen, etc.)
  3. 3 Select antibodies with confirmed reactivity in native protein state
  4. 4 Consider alternative antibodies specifically validated for IHC with your tissue type
  5. 5 Test antibody in pilot experiment with known positive control tissue
Related Video (3)
Cell Signaling Technology ★ 85
Tissue Controls & More for Immunohistochemistry | CST Tech Tips
"Directly addresses antibody validation and control strategies (positive/negative/isotype controls) essential for diagnosing when antibodies fail in IHC"
Bilibili (China-Accessible Mirrors) ★ 78
Immunohistochemistry on Paraffin-Embedded Sections (CST Demo)
"CST expert-led protocol covering antigen retrieval and native epitope preservation in paraffin sections, directly relevant to antibody-tissue conformation mismatch"
Cell Signaling Technology ★ 72
Better IHC Step 1: Antigen Retrieval
"Focused optimization series on antigen retrieval techniques that improve epitope accessibility, addressing the core issue of native vs. denatured protein recognition"
Source: abcam.com ↗
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