Immunohistochemistry (IHC) Tips

Cell permeabilization:

For a better penetration of antibody, Triton X-100 or protease K is commonly used.

Triton X-100 can dissolve the lipid on cell membranes, nuclear membranes and organellar membranes, therefore big molecules such as antibodies can get into cytoplasm and nucleus to bind antigens. In immunohistochemistry (>10μm sections) and immunocytochemistry, as a detergent, Triton X-100 works with concentration of 0.5%-1% for immunohistochemistry and 0.05% for immunocytochemistry in PBS buffer.

*As for paraffin section with thickness of 4μm, cell permeabilization is not necessary.