Home › Failure Case Library › False negative — no staining at all
Immunohistochemistry (IHC) critical

False negative — no staining at all

Symptom
Even the positive control shows no staining; expected positive regions are completely blank; the experiment failed.
Common Causes
  1. 1 Antibody is degraded or stored incorrectly
  2. 2 Antigen retrieval failed
  3. 3 Detection reagent has expired
  4. 4 Protocol steps were missed or done in wrong order
Solutions
  1. 1 Use a fresh antibody with proper storage (aliquot, -20 °C)
  2. 2 Re-optimize antigen retrieval conditions (try a different buffer or longer time)
  3. 3 Check expiry dates of all reagents (DAB, secondary antibody, blocking serum)
  4. 4 Strictly follow the protocol step by step; include a positive control slide every time
Related Video (3)
Cell Signaling Technology ★ 72
Immunohistochemistry Protocol for Paraffin embedded Tissue Sections
"Full CST IHC protocol covering antibody handling and proper technique, useful for preventing degraded-antibody failures."
Cell Signaling Technology ★ 85
Tissue Controls & More for Immunohistochemistry | CST Tech Tips
"Directly addresses positive/negative tissue controls, which is exactly how this total-failure false negative is diagnosed."
Bilibili (China-Accessible Mirrors) ★ 68
Immunohistochemistry on Paraffin-Embedded Sections (CST Demo)
"Step-by-step paraffin IHC demonstration showing correct reagent/antibody use to perform the technique properly."
Source: xiaohongshu.com ↗
← Back to all cases