Knowledge Base › Troubleshooting Atlas › IHC
Over-digestion during antigen retrieval
Also known as: Antigen retrieval artifact
Tissue sections appear overly pale or washed out, with loss of detail in cellular morphology. Antigens may be poorly accessible or non-detectable.
Where it usually appears: Typically observed during the immunohistochemistry (IHC) staining process after excessive heat or enzyme exposure during antigen retrieval.
- Prolonged exposure to heat or enzyme during retrieval
- Incorrect retrieval buffer concentration
- Inappropriate retrieval temperature
- Insufficient cooling time after retrieval
- Check temperature settings on water bath or microwave
- Verify retrieval buffer composition and concentration
- Inspect tissue morphology under a microscope
- Adjust retrieval time and temperature in future runs
- Optimize buffer concentration based on tissue type
- Re-evaluate enzyme digestion time
- Establish standard retrieval protocols with time and temperature guidelines
- Regularly calibrate equipment used for antigen retrieval
- Train staff on proper retrieval techniques
This atlas is for educational purposes for laboratory professionals. It does not replace your institutional SOPs, manufacturer instructions, validated protocols or pathologist judgment. Adapt any action to your laboratory and equipment.
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