Antibodies that label paraffin-embedded mouse tissues: a collaborative endeavor.
Biological-Markers, Cell-Membrane, Cell-Nucleus, Cytoplasm, Epitope-Mapping, Epitopes, Female, Fixatives, Fluorescent-Antibody-Technique-Indirect, Immunoenzyme-Techniques, Male, Mice, Mice-Inbred-C57BL, Paraffin-Embedding, SUPPORT-U-S-GOVT-P-H-S, Viral-Proteins
see Reprint Collection
Toxicol Pathol 2004 Mar-Apr; 32(2):181-91.
Histology and immunohistochemistry are important tools in the study of human diseases and their respective animal models. The study of mouse models has been hampered by the absence of a large set of mouse-specific antibodies adapted to paraffin-embedded tissues. A total of 196 antibodies were tested on paraffin-embedded mouse tissues preserved in five different fixatives (Fekete's acid-alcohol-formalin, 10% neutral buffered formalin, 4% paraformaldehyde, IHC Zinc Fixative, and Bouin's fixative). The antibodies were targeted to proteins of the cytoplasm (n = 100), plasma membrane (n = 48), nucleus (n = 36), extracellular compartment (n = 5), cytoplasm/cell membrane (n = 4), and viral proteins (n = 3). A total of 83 antibodies provided an adequate signal to noise ratio. Of these, adequate labeling required heat-mediated epitope retrieval or enzymatic digestion for 32 and 8 antibodies, respectively. Epitope recognition was best for tissues fixed with Fekete's acid-alcohol-formalin. However, some proteins could be detected only in IHC Zinc Fixative, confirming that there is no single fixative suitable for the preservation of all epitopes. Four of 13 antibodies that failed to label their cellular targets on tissue sections successfully labeled whole-mount tissues, indicating that tissue processing plays an important role in epitope degradation. Regularly updated information on immunohistochemistry of normal and neoplastic mouse tissues is accessible online at (http://tumor.informatics.jax.org); links to antibody suppliers' web sites are provided.
Mikaelian, I; Nanney, L B.; Parman, K S.; Kusewitt, D F.; Ward, J M.; Naf, D; Krupke, D M.; Eppig, J T.; Bult, C J.; Seymour, R; Ichiki, T; and Sundberg, J P., " Antibodies that label paraffin-embedded mouse tissues: a collaborative endeavor." (2004). Faculty Research 2000 - 2009. 740.