Target protein absent or below detection threshold in tissue

Symptom

No staining observed in experimental tissue samples. The target protein may not be expressed in the tissue type or developmental stage being examined, or present at undetectable levels.

1. 1 Target protein not expressed in the specific tissue type examined
2. 2 Protein expression below detection limit of the IHC method
3. 3 Developmental stage or physiological condition not supporting protein expression
4. 4 Tissue heterogeneity with target protein present only in specific cell populations

1. 1 Review scientific literature to confirm protein expression in your tissue type
2. 2 Use known positive control tissue verified to express target protein
3. 3 Validate protein presence by western blot or RT-PCR before IHC
4. 4 For low-abundance proteins, implement signal amplification (biotin-streptavidin, TSA)
5. 5 Consider tissue from different developmental stages or treatment conditions
6. 6 Examine multiple tissue sections to account for spatial heterogeneity

Related Video (3)

Cell Signaling Technology ★ 90

Tissue Controls & More for Immunohistochemistry | CST Tech Tips

"Directly addresses how to verify if antibody is working and use tissue controls to distinguish true absence from technical failure."

Cell Signaling Technology ★ 75

Immunohistochemistry Protocol for Paraffin embedded Tissue Sections

"Provides a correct IHC protocol on paraffin sections, useful to rule out procedural errors causing no staining."

Bilibili (China-Accessible Mirrors) ★ 70

Immunohistochemistry on Paraffin-Embedded Sections (CST Demo)

"Shows a step-by-step CST protocol for paraffin sections, helpful for checking technique if target is known to be present."