Table 5.

Recommendation on the details to be included in the methods in published manuscripts with immunohistochemistry results.

Catalog number and lot number of the antibody, company, monoclonal/polyclonal

Species specificity

Evidence supporting the specificity of antibody; it is not just enough to quote a manuscript which uses that antibody. The reference should be to a publication in which different tools are used to prove the specificity of an antibody

Negative controls used; should include staining with preimmune serum or isotype specific immunoglobulin; omitting primary antibody is not a sufficient control; comments on positive controls used; comments on internal positive and negative controls (when possible); adsorption controls should not be considered as proof of specificity by itself

Fixative, duration of fixation, duration in alcohol (if held in between fixation and processing)

Antigen retrieval method, buffer used for antigen retrieval, pH and molarity of the buffer,

Antibody dilution along with protein concentration of the antibodies used

Duration of incubation with antibody; the use of “overnight incubation” should be discouraged unless it is qualified by approximate number of hours.

Detection method used