Immunohistochemistry
Background: immunohistochemistry
consists of the following steps:
- Primary antibody binds to specific antigen.
- Antibody-antigen complex is bound by a secondary, enzyme-conjugated,
antibody. To amplify the staining, we use a complex of peroxidase-antiperoxidase
(PAP), avidin-biotin-peroxidase (ABC) or avidin-biotin alkaline phosphatase.
- In the presence of substrate and chromogen, the enzyme forms a colored
deposit at the sites of antibody-antigen binding. The color of the
deposit depends on the chromogen used (see figures
below).
Suitable for : all tissues,
provided antigenicity is not compromised by fixation.
Not suitable for : detection
of those antigens that require the use of either special fixatives or
frozen tissue. Contact us for more information.
Note : Histology and Tissue
Processing offers a service to try out new and custom antibodies.
Antigens routinely detected by immunohistochemistry:
| Cellular Mechanism |
Antigens (partial listing only, contact
us about other antigens). |
| Proliferation |
BrdU (bromodeoxyuridine), Ki-67, PCNA
(Proliferating Cell Nuclear Antigen) |
| Cell Cycle |
Cyclin D1, E2F1, E2F4, Rb, p21, p27 |
| Differentiation |
Keratins 1, 5, 6, 8, 10, 13, 14, 19;
CD31 |
| Tumorigenesis |
p53, Rb, c-myc |
Immunoperoxidase staining of (left) keratin 5 in formalin-fixed
mouse skin, and (right) p53 in formalin-fixed, benzo[a]pyrene-treated
hamster cheek pouch.
Alkaline phosphatase staining of keratin 5 (red) in formalin-fixed mouse mammary gland. The brown nuclei are TUNEL-positive (apoptotic).
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