Anti-Rabbit Secondary Antibodies
Anti-Rabbit Secondary Antibodies are antibodies that are used to detect and visualize the presence of primary antibodies that are raised in rabbits. They are generated by immunizing a host animal with a pooled population of immunoglobulins (Ig) from the target species, in this case, rabbits, and then collecting and purifying the resulting antibodies. Anti-Rabbit Secondary Antibodies are available in host species including goat, donkey, mouse, chicken, and sheep, and come in a variety of conjugates, including unlabeled antibodies, enzyme conjugates (alkaline phosphatase (AP) or horseradish peroxidase (HRP)), and biotin. They are also available as whole IgG.
Choosing the right anti-rabbit secondary antibody for an experiment depends on several factors, including the application, host species of the primary antibody, isotype of the secondary antibody, conjugate type, and pre-adsorption or cross-adsorption. The following steps can help in selecting the right antibody:
- Determine the host species of the primary antibody: The secondary antibody should be raised against the host species used to generate the primary antibody. For example, if the primary antibody is raised in rabbit, an anti-rabbit secondary antibody raised in a different species other than rabbit, such as goat, donkey, or mouse, should be used.
- Determine the isotype of the primary antibody: The secondary antibody should be directed against the class/isotype of the primary antibody. For instance, if the primary antibody is a mouse IgG1, an anti-mouse IgG or a less specific F(ab) fragment anti-mouse IgG should be used.
- Choose the conjugate type: Secondary antibodies can be enzyme-labeled (peroxidase, alkaline phosphatase), fluorochrome-labeled (FITC, R-PE, Alexa-Fluor®), or biotinylated. The choice of conjugate type depends on the application.
- Consider pre-adsorption or cross-adsorption: Pre-adsorption or cross-adsorption can help reduce non-specific binding. For example, if the primary antibody is raised in rabbit, an anti-rabbit secondary antibody that has been pre-adsorbed against other species, such as mouse or human, can be used.
- Check the compatibility with the assay: The secondary antibody should be compatible with the assay being used, such as Western blot or immunohistochemistry.
