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How to Select an Antibody for Target Protein?

There are always more than one antibody can be chosen for detection of any target protein. Here are several tips for narrowing the selection range and choosing the appropriate antibody for your experiment.

Refer to the literatures and publications

Go through the published researches and check that which antibodies targeting your protein of interest have been successfully used by others for similar applications. Keep them at the top of your list and make the best choices between them.

Epitope of the antibody

Select the one which is raised against an immunogen that is identical to or covers the interested part of your protein.

Applications of the antibody

For example, western blots work on denatured proteins, while flow cytometry is compatible with native configuration proteins. Some antibodies apply to the experiments including western blots (WB), Immunofluorescence (IF), and immunohistochemistry (IHC), but others may only apply to one or two of them.

The specificity species of the antibody and samples

The antibody which works well in one cell type may be not always suitable for your cell type. What’s more, if an antibody was raised in the same species as your target one, it may cause problems later at the detection stage.

Monoclonal antibodies or polyclonal antibodies?

Monoclonal antibodies often provide a high level of specificity, low non-specific binding, and low background signals because they recognize a single epitope on a target protein, while polyclonal antibodies tend to be less specific because they are raised against multiple epitopes on your protein. However, polyclonal antibodies can be more effective at detecting a target even if a few epitopes are masked by cross-linking. Monoclonal antibodies are good for flow cytometry where multiple epitope-recognition by polyclonal antibodies would lead to inaccurate data. Polyclonal antibodies are generally suitable for applications like IP where multiple epitope recognition is desirable.