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Immunohistochemical (IHC) antibodies are used to tag antigens in scientific studies. They have specificity and binding ability and can be used in staining and precipitation techniques. Primary and secondary antibodies are commonly used, with the latter being labeled with identifying molecules.
An immunohistochemical (IHC) antibody is a commonly used reagent in scientific studies to tag the antigens of interest. An antigen is any small molecule that can cause an immune reaction that forces the production of antibodies against that single antigen. Based on the mammalian immune system, the production of antibodies is a single step in a process of identifying material as foreign so that the body can then attack and expel the invasive substance, whether it is a virus, bacterial infection, or a small molecule like pollen.
Antibodies can have tremendous specificity and binding ability in labeling antigens. This specificity is exploited to the advantage of scientists when antibody production is geared towards identifying specific antigens of interest. A thin section of tissue can be stained with an antibody so that the investigator can easily identify all antigenic sites present in that sample. Many companies specialize in the creation of antibodies, making a vast collective catalog of antibodies available to scientists and investigators.
An IHC antibody is used in several experimental techniques. Staining of antibodies in thin sections of tissue embedded in paraffin or plastic is often used so that the antigens can be found in situ. An IHC antibody can also be used in experimental precipitation techniques such as a co-immunoprecipitation assay, focusing on bringing antigens out of solution to quantify the amount of antigen present in a large piece of tissue. Assays that measure the amount of light that is able to pass well through a tissue culture plate after a supernatant has been exposed to the bottom of a well pre-coated IHC antibody are often used to quantify the amount of an antigen in a tissue culture plate. homogenized tissue sample.
It is possible to have antibody staining protocols that rely on the use of a single antibody for antigen detection. These protocols are usually used on tissue samples embedded in a non-reactive substrate and mounted on a glass slide. These single antibody staining procedures are also popular in laboratories that label their antibodies with sensing molecules such as fluorophores.
More commonly, however, antibody staining protocols require the use of primary and secondary antibodies. A primary IHC antibody is raised against a specific antigen, which may not commonly be studied. A secondary IHC antibody will be bred against the primary IHC antibody. However, the secondary antibody will be labeled with an identifying molecule such as biotin or a fluorophore. This two-antibody technique is common because companies save money by adding fluorophores or expensive labeling reagents to large quantities of antibodies raised against immune markers of a certain species rather than labeling expensive reagents to an antibody raised against an antigen that could be studied by very few scientists.
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