Immunohistochemistry (IHC) and Immunofluorescence (IMF) are essential pathology tools for the diagnosis and treatment of many common pathological conditions. The ITPC has developed a number of standard IHC and IMF protocols for many antibodies. Some of the common IHC staining protocols include BrdU, ED2, F480, and GSTp. Additional immunohistolochemical markers can be performed. Paraffin embedded tissue sections to be stained or blocks should be brought to the ITPC Lab. The ITPC will assist investigators in the establishment of IHC protocols for antibodies of interest. Antibodies should be provided by the investigator along with related literature references. The ITPC also offers evaluation of the tissue section / IHC results.  Assistance is provided in antibody titration and interpretation of the stain.

Specimen samples: Ideally prior to tissue specimen collection, the investigator should discuss the protocol with ITPC staff for optimal fixation and processing procedures for the target molecule of interest.

Routine Tissue Preparation for Immunohistochemistry and Immunofluorescence:

Tissues should be fixed overnight in 10% neutral buffered formalin (roughly 22-24 hours) or 3% Paraformaldehyde (see below)

Then transfer to 70% ethanol

The tissue is ready to be shipped to the histology lab

Tissue can stay in 70% ethanol for two weeks before changing

3% Paraformaldehyde:
535ml distilled H20
80 ml 10X PBS
185 ml Paraformaldehyde* (Ultrapure or EM grade)

*Purchase the paraformaldehyde from Sigma or EM Sciences only.