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.

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

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 ★ 88

Tissue Controls & More for Immunohistochemistry | CST Tech Tips

"Directly addresses validating antibody performance in IHC using positive/negative tissue and isotype controls, the core of this failure."

Bilibili (China-Accessible Mirrors) ★ 72

Immunohistochemistry on Paraffin-Embedded Sections (CST Demo)

"CST paraffin IHC demo covering antigen retrieval and protocol steps where antibody-specificity failures are diagnosed."

Cell Signaling Technology ★ 65

Better IHC Step 1: Antigen Retrieval

"Explains antigen retrieval, a key variable when antibodies fail to stain native/non-retrieved epitopes in tissue sections."