Nuclear protein not detected due to insufficient permeabilization (Immunohistochemistry (No Staining))

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Accepted Answer

No staining observed when targeting nuclear proteins. Antibody cannot access nuclear compartment despite proper antibody concentration and incubation time. Common causes: 1) Antibody cannot penetrate nuclear membrane due to insufficient permeabilization 2) Weak or absent permeabilizing agents in blocking and antibody buffers 3) Nuclear envelope remains intact preventing antibody access to nuclear epitopes Solutions: 1) Add strong permeabilizing agent like Triton X to blocking buffer 2) Include Triton X in antibody dilution buffer 3) Increase permeabilization time for better nuclear penetration 4) Use concentration of 0.1-0.5% Triton X-100 for nuclear targets