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Secondary Antibodies

Secondary antibodies can be monoclonal or polyclonal and are useful for detecting, sorting, and purifying antigens. A typical secondary antibody is a polyclonal antibody labelled with a probe which binds to a primary antibody which in turn has been bound to an antigen of interest. Because multiple secondary antibodies can bind to a single primary antibody it is possible to achieve increased sensitivity due to signal amplification.

Enzo Life Sciences offers secondary antibodies from a variety of species featuring conjugates appropriate for ELISA, WB, IHC, ICC, and FC.


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Target Ig Class

Specificity of Secondary Antibodies

A secondary antibody is created by immunizing a host animal with an antibody from an animal of a different species. Common secondary antibody hosts are goat, donkey, sheep, chicken, and rabbit.

It is possible to create specificity to whole Immunoglobulins or smaller fragments depending on what is used to immunize the host animal and through downstream purifications. Most secondary antibodies are generated against a pool of IgG from the target species. This will allow binding to all Ig subclasses, heavy and light chains, fragments, and conserved domains. If a more specific target is desired, the host can be immunized with a specific target, for example IgG2 only. This will result in a secondary that only binds to species specific IgG2. Specificity can also be increased via column purification through a matrix to adsorb out cross-reacting antibodies.

Considerations for Selecting a Secondary Antibody

  1. Host of the primary antibody
  2. Class/subclass of the primary antibody.
  3. Cross reactivity of the secondary.
  4. Preferred label based on downstream detection method.

Popular Applications for Secondary Antibodies

  • Western blot
  • Immunohistochemistry
  • Immunocytochemistry

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