Immunoglobulin fluorescein method

The following procedure is a suggested method for using NHS-fluorescein to label immunoglobulins. 

Acceptable bridging molecule systems have been developed which have also simplified the utilization of different detection systems. To illustrate this point, a researcher who has developed a unique monoclonal antibody (a primary antibody) in the mouse may select from a variety of commercially available products consisting of different detection systems (e.g. fluorescein, alkaline phosphatase, colloidal gold) attached to an immunoglobulin that will specifically bind to mouse antibodies (a secondary antibody). In this way the researcher may readily obtain and test a number of detection methods for visualizing target-probe interactions without having to directly label the monoclonal antibody probe. For nucleic acid probes, which in themselves are not readily immunodetectable, it is useful to incorporate or attach detectable moieties to the nucleotides. 

When used to detect mouse monoclonal antibodies this secondaiy antibody would have anti-mouse immunoglobulin specificity. The method can be adapted for a number of reporter molecules, for example other enzyme labels such as alkaline phosphatase, and fluorescent molecules such as fluorescein isothiocyanate (FITC). 

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