By in General FAQs

How to choose suitable Secondary antibody?

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  • Which animal was used to generate your primary antibody? For instance, if your primary antibody is raised in a rabbit, you need to choose an anti-rabbit secondary antibody; if it is raised in a mouse, you need to choose an anti-mouse secondary antibody.
  • What class (or subclass) is your primary antibody? The secondary antibody should match the class/subclass of the primary antibody. This is most important for monoclonal antibodies because different classes (or subclasses) can be found. Since polyclonal antibodies, such as those raised in rabbit, are usually of the IgG class, anti-IgG secondary antibodies are commonly used.
  • What application do you want to perform? Enzyme-linked secondary antibodies such as those linked to peroxidase or alkaline phosphatase, are commonly used for ELISAs, Immunoblotting and Immunohistochemistry applications. For Immunofluorescence and Flow Cytometry, which rely on fluorescence, a fluorochrome-conjugated secondary antibody is needed.