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Immunohistochemistry (No Staining) moderate

FFPE tissue shows no staining from incomplete deparaffinization

Symptom
Paraffin-embedded tissue sections show no immunostaining. Residual paraffin creates a physical barrier preventing antibody access to tissue antigens.
Common Causes
  1. 1 Insufficient deparaffinization time leaving paraffin residue in tissue
  2. 2 Old or degraded xylene with reduced solvent effectiveness
  3. 3 Incomplete paraffin removal blocking antibody penetration into tissue sections
Solutions
  1. 1 Deparaffinize sections for longer duration
  2. 2 Use fresh xylene for optimal paraffin removal
  3. 3 Perform additional xylene incubation steps (3 changes of 5 min each)
  4. 4 Verify complete deparaffinization by checking tissue appearance before proceeding
Related Video (3)
Bilibili (China-Accessible Mirrors) ★ 95
Immunohistochemistry on Paraffin-Embedded Sections (CST Demo)
"Directly demonstrates IHC on paraffin-embedded sections with explicit focus on antigen retrieval and deparaffinization steps, addressing the root cause of incomplete deparaffinization."
Cell Signaling Technology ★ 85
Immunohistochemistry Protocol for Paraffin embedded Tissue Sections
"Comprehensive IHC protocol for paraffin-embedded tissue from Cell Signaling Technology covers complete deparaffinization workflow essential for preventing no-staining failures."
Cell Signaling Technology ★ 78
Better IHC Step 1: Antigen Retrieval
"Focused troubleshooting series on antigen retrieval as Step 1, directly relevant to understanding how improper deparaffinization leads to poor staining outcomes."
Source: abcam.com ↗
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