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Immunoflourescense

Immunoflourescence is a technique that utilizes antibodies linked to fluorescent molecules to show the location of a particular protein in a cell or tissue. The most commonly used fluorescent tags are fluorescein (emits green light), and rhodamine (emits red light). Multiple differently-tagged antibodies can be used to mark multiple targets in a given cell. One common way to increase the signal is to first use a nonlabelled antibody which can be subsequently recognized by multiple fluorescent-tagged anti-antibody antibodies (secondary antibodies). One risk of this type of signal amplification is that the secondary antibodies may bind generically to other proteins in the cell (or to other antibodies) and thus give a nonspecific signal. In order to control for this, the specificity of the antibodies used should be verified by performing a Western blot and testing that the antibodies only mark a single band.

Editor(s): John Bracht

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