Rodent Immunohistochemistry
Rat and Mouse Protocols
Sources of Antibodies
(All sites listed are non-federal Web sites.)
References of Antibodies
Examples of Antigen Expression
Illustrated on this page and related pages are examples of antigens we have worked with. Some have been difficult to demonstrate. For these, we explain the tricks to use for demonstrating these antigens. If you have a specific problem in immunostaining involving rodents, you can e-mail Dr. Ward for his comments.
Nuclear Antigens
p53 nuclear reactivity in human cell line with p53 mutation. Note variation in nuclear immunoreactivity, paraformaldehyde fixation
T/ebp (TTF-1) in bronchiolar epithelial cells, requires Carnoy's fixation. Courtesy of Dr. Minoo, USC.
BrdU in basal cells of mouse skin after TPA exposure.
PCNA in mouse liver preneoplastic focus.
Apoptosis can be identified with H&E stained sections (left) or apoptosis kits (right). Both methods illustrate the same apoptotic bodies in the mouse stomach.
![Apoptosi identified apoptosis kits photograph](https://webarchive.library.unt.edu/eot2008/20081012203214im_/http://www3.niaid.nih.gov/NR/rdonlyres/2A747F5E-93E8-4A85-AFC1-E64B0FE20C91/0/Apoptosisapop2.jpg)
Cytoplasmic Antigens
Left - Actin in myoepithelial cell. Formalin fixation. Slide courtesy of Dr. Sabine Rehm. Middle - Keratin (high molecular weight) in bile duct epithelium and oval cells of mouse liver. Ethanol is often the best fixative to use to demonstrate keratins. Right - IgG in plasma cells in mouse lymph node. Immunoglobulins are best demonstrated with Bouin's, B-5 or paraformaldehyde fixation.
![plasma](https://webarchive.library.unt.edu/eot2008/20081012203214im_/http://www3.niaid.nih.gov/NR/rdonlyres/0B6B2F77-1966-44A1-8391-328C8CC31F21/0/plasma.jpg)
Left - GSTP in human liver bile duct, ductule and oval cells. Formalin. Note similarity to keratin in similar mouse liver lesions. Right - ras p21 (in a granular cytoplasmic staining pattern) in normal rat renal tubule cells with a unique monoclonal antibody. Contrast with membrane staining seen below in tumor cells. Bouin's fixation.
![ras p21](https://webarchive.library.unt.edu/eot2008/20081012203214im_/http://www3.niaid.nih.gov/NR/rdonlyres/31D0FA89-FE92-4738-BB24-62C76CE544E0/0/p21k.jpg)
Antigen in endothelium of venules and capillaries in a mouse transplantable tumor
![tumor vessels photograph](https://webarchive.library.unt.edu/eot2008/20081012203214im_/http://www3.niaid.nih.gov/NR/rdonlyres/0BE7DA13-E06B-41A5-9326-26FA77C2E567/0/tumorvessels.jpg)
Positive Cell Controls For Immunohistochemistry
The Pellet Technique: In Vitro Cell Pellets As Controls For Immunohistochemistry
- Scrape bottom of flask with rubber policeman
- Centrifuge media and cells to produce pellets
- Fix in formalin, Bouin's, frozen etc.
- Embed in paraffin
- Perform Immunohistochemistry for antigen
- Compare with tissue sections
Left - Pellets in tubes after centrifugation of cell lines. Middle - ras p21 on cell membranes of cultured cells prepared by pellet technique as shown on left. Cell line with 30 copies of H-ras. Bouin's fixation. Right - an antigen immunoreacting on cell membranes and in Golgi area of cells.
![antigen immunoreacting](https://webarchive.library.unt.edu/eot2008/20081012203214im_/http://www3.niaid.nih.gov/NR/rdonlyres/27903D60-D983-4964-80E0-D7F6D338D3B7/0/pelletkai1b.jpg)
Mouse Hematopoietic Pathology
See books in Pathology of Genetically Engineered Mice
Atlas of Mouse Hematopathology (non-federal Web site)
back to top