High background
Unexpected staining patterns can commonly be due to suboptimal digestion conditions. Use examples below to determine possible issue and solution.
Nuclear Background
Standard pretreatment (15 min target retrieval + 30 min protease) on Mm kidney
Problem
Pretreatment conditions not optimal
- Tissue is over-digested
- Tissue is under-digested
Solution
Optimize pretreatment conditions:
- Decrease boiling and/or protease if tissue is over-digested
- Increase boiling and/or protease if tissue is under-digested
