Superclonal secondary antibodies represent a breakthrough in recombinant antibody technology. By increasing the number of different species of serum components in the adsorption column, one can generate antibodies that exhibit cross-reactivity to fewer and fewer species.įor even further elimination of cross-reactivity, try Invitrogen™ Superclonal™ secondary antibodies. The secondary antibodies that recognize these other species’ antibodies or serum proteins will bind to the column, whereas those without cross-reactivity to flow through. Cross-adsorption involves passing the affinity-purified secondary antibody over a column containing immobilized antibodies or serum proteins from other species. Several kinds of anti-mouse and anti-rabbit secondary antibodies from other host species are also available.Ĭross-adsorption of secondary antibodies is helpful in eliminating cross-reactivity from other nontarget antibodies and proteins, thereby increasing the antibody’s specificity. Consequently, goat secondary antibodies against mouse IgG and rabbit IgG are commercially available in the widest variety of forms. Goat is the host species most easily and frequently used by manufacturers to produce polyclonal anti-mouse and anti-rabbit secondary antibodies. Most primary antibodies are produced in mouse or rabbit hosts therefore, anti-mouse IgG and anti-rabbit IgG are the most popular types of secondary antibodies. For instance, if a primary antibody was derived from a rabbit, then one must use an anti-rabbit secondary antibody that was generated in a species other than rabbit. Keep in mind that the species used to generate the secondary antibody should always be different from the primary antibody host and target species. We offer secondary antibodies generated in chicken, donkey, goat, mouse, rabbit, rat, and sheep. For example, when rabbit IgG is used to immunize a goat, the result is production of goat anti-rabbit secondary antibodies. The host species is the animal in which the secondary antibody was generated. Conjugation (which is amino acid-specific) can interfere with the primary antibody’s recognition of the antigen. By using secondary antibodies, one avoids needing to chemically label (conjugate) primary antibodies, which are more specialized and costly to obtain. For example, the same primary can be used with a secondary antibody conjugated to HRP in a western blot and with a different secondary antibody conjugated to Alexa Fluor 488 dye in immunofluorescence.Īlso, secondary antibodies enhance detection by localizing more conjugate at the antigen than is possible with a labeled primary antibody. Using a secondary antibody makes it easy to choose a different conjugate no matter the primary antibody. It is possible to use conjugated primary antibodies, but secondary antibodies provide many advantages. Many secondary antibodies are conjugated to other molecules, like Alexa Fluor dyes or horseradish peroxidase (HRP), which enable detection of the secondary antibody. Secondary antibodies are often used in combination with primary antibodies to detect target proteins in various immunoassays, like western blots, ELISA, and immunofluorescence. A secondary antibody is an antibody designed to target a primary antibody.
0 Comments
Leave a Reply. |
AuthorWrite something about yourself. No need to be fancy, just an overview. ArchivesCategories |