Immunocytochemistry (ICC) — Step-by-Step Protocol (Bio-Techne)

severe

Epitope Degradation from Harsh Permeabilization

Loss of antibody binding or weak signal for intracellular targets despite successful cell permeabilization. Positive controls show reduced or absent staining.

💡 4 · ✓ 4

severe

Incorrect Permeabilization Method for Target Location

Weak or absent staining of nuclear transcription factors when using detergent-based permeabilization, or poor cytoplasmic protein detection with alcohol-based methods.

💡 4 · ✓ 4

severe

Failed Intracellular Antigen Detection After Fixation

Intracellular markers such as cytokines, transcription factors, or phosphoproteins show no signal despite proper antibody validation. Surface markers stain normally, but internal targets remain undetected indicating permeabilization issues.

💡 4 · ✓ 5