Secondary antibodies are bound to the primary antibodies and help in identifying, categorizing and cleansing of objectified antigens. Secondary antibody is exclusively adept in immunolabeling which is a process that detects antigens in specific tissues, cells and organs. Secondary antibodies are appended to the primary antibodies which are precisely appended to the antigens that are targeted.
In the process of immunolabeling, the Fab Domain of the secondary antibody appends to pollen or antigen and discloses its FC domain to the secondary antibodies. Then, the Fab domain of the secondary antibody appends with the Fc domain of the primary antibody. As the Fc domain is continual inside the same animal section, only a single type of secondary antibody is needed to append to several kinds of primary antibodies. This curtails the cost by having to label on a single kind of secondary antibody, instead of having to label several kinds of primary antibodies. Due to numerous secondary antibodies appending to a primary antibody, the secondary antibody assist in boosting the signal and sensitivity.
Functions of Secondary Antibodies
Secondary antibodies can adjoined to various enzymes such as rhodamine derivatives, alkaline phosphatase, horseradish peroxidase or other molecules that can be useful in several applications. Secondary antibodies are used in several analysessuch as:
- ELISA- is a test that makes the use of antibodies and change of color in order to detect a substance including tests of HIV.
- Immunocytochemistry- is a regular laboratory method that is used in order to visualize and localize a definite pollen or protein present in the cells by making the use of a definite primary antibody that appends to it.
- The Western Bolt- also known as the protein immunobolt is a broadly used method of analysis specifically in molecular biology to identify definite proteins in tissue extracts.
- Immunostaining- is a common tern that administers to any kind method that is based on antibodies to identify a definite protein in cells or tissues.
Secondary antibodies are brought out by inoculating a host animal that has antibodies from another species. For example, secondary antibodies that are anti-cow are increased by injecting cow antibodies into an animal different than a cow.To generate secondary antibodies, animals like rabbit, got and donkeys are usually used as host species however, other species can also be useful. Inoculating a goat with purified cow immunoglobulins will produce goat anti-cow immunoglobulins antibodies and will append to all categories of cow immunoglobulins and also with any other molecule that has domain in common. Furthermore, the purification of secondary antibodies can be done by putting it through columns that contain disabled proteins different than those that are being used to inoculate the host.