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Secondary antibodies are commonly used as detection reagents in many different types of immunoassays such as western blotting, immunohistochemistry and immunocytochemistry or cellular imaging. However, the importance of selecting the right secondary antibody is sometimes overlooked.
Secondary antibodies, like any other antibody, are designed to target a specific antigen. All secondary antibodies target immunoglobulins (Igs) so they can bind primary antibodies and serve as detection reagents in immunoassays. Thus, many of the factors that require consideration when selecting the appropriate primary antibody for your experiment, also apply to secondary antibodies. Selecting the right secondary for your assays can have a significant positive impact on your results. However, a poor secondary can have as much of a detrimental effect on your experiment as a poor performing primary antibody. Three parameters are mainly used to determine the performance of antibodies:
Performance of secondary antibodies is influenced by many different factors, for which the researcher can normally select from a range of different alternatives in order to ensure optimal results:
|Target Protein (Igs)|
|Isotype||In particular situations it may be good to consider the isotype of the secondary antibody itself. For instance, antibodies raised in mice which isotype is IgG2a will strongly bind to human leukocytes.|
|Clonality||The benefits and limitations of clonality for primaries also apply to secondaries.|
Secondaries you can trust
We have generated a high quality range of well-defined secondaries to ensure the success of your experiments. Each of these secondaries has been extensively validated in key applications and quantitative data on their performance is available on their datasheets.
Our catalogue also includes a broad range of secondaries to ensure you find the product you need such as fragments or subclass-specific secondaries among many others.
Alexa Fluor® is a registered trademark of Life Technologies. Alexa Fluor® dye conjugates contain(s) technology licensed to Abcam by Life Technologies.