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Simple, Sensitive Isotyping of Mouse Monoclonal Antibodies

Monoclonal antibody isotyping in only 5 minutes

Weiping Yang Matt C. Petre Darrel Holmes Peter L. Pingerelli
Stratagene Cloning Systems, Inc.

Stratagenes IsoDetect mouse monoclonal antibody isotyping kit is a simple, rapid and sensitive method for identifying mouse monoclonal antibody isotypes in a single step. Each IsoDetect test card is a self-contained assay, using colloidal gold-conjugated anti-mouse antibodies for isotype determination of 0.1 g/ml of antibody in 5 minutes.

For decades, antibodies have been used as key reagents for biological research. The development of hybridoma technology, which generates monoclonal antibodies, has resulted in the development of a significant number of immunoglobulins for research and diagnostics. As knowledge concerning the production of monoclonal antibodies has grown, these powerful tools have become more commonplace, and thousands of new monoclonal antibodies are made each year.

Mouse immunoglobulins have been classified into five primary classes (IgG, IgM, IgA, IgD and IgE) according to differences in their heavy chain polypeptides (the immunoglobulin classes have g, , a, d and e heavy chains, respectively). The antigenic differences among the five classes are located primarily on the Fc fragments of the heavy chains. These differences allow immunoglobulins to function at particular stages in a variety of immune responses. IgG molecules are further subclassed based on the antigenicity of the four isotypes. Mouse IgG isotypes are designated IgG1, IgG2a, IgG2b and IgG3.

Because the biological functions and biochemical characteristics of classes and isotypes differ, distinguishing the classes and isotypes of an immunoglobulin molecule is critical. Knowl
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