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Simple, Sensitive Isotyping of Mouse Monoclonal Antibodies

e sample. This assay is highly sensitive and relatively fast (2 to 4 hours), but the required multistep operations of the assay are labor-intensive. A plate reader, which is best for quantitating ELISA results, may be an investment that is prohibitive for some laboratories.

A rapid method of isotyping uses anti-mouse antibody that is conjugated to an indicator molecule, such as colloidal gold or latex, and isotyping antibodies coated on a membrane in separate, discreet lines. The antibody in a sample forms an antigen-antibody complex with the indicator conjugate and is exposed to the isotyping membrane. When the sample complex meets the specific isotyping antibody, a colored line appears. Results are judged by reading the line at a specific location. This format allows multiple samples to be analyzed in a simple one-step operation. Fast results (5 minutes) and high sensitivity (as low as 1.0 g/ml of antibody, according to manufacturers) characterize this newest immunoglobulin isotyping method. Because this method is easy to use and gives rapid results, it is likely to replace the ELISA method for mouse immunoglobulin isotyping.

Single-Step Isotyping

figure 1

Stratagenes IsoDetect mouse monoclonal antibody isotyping kit is a rapid method for determining the isotypes of mouse monoclonal antibodies. This kit uses a colloidal-gold conjugation technique to label rabbit anti-mouse l and k light chain antibodies as indicating reagents, and rabbit anti-mouse isotyping immunoglobulin coated on a membrane. When a specific mouse immunoglobulin is present in a sample, a complex containing mouse immunoglobulin and rabbit anti-mouse l or k gold-conj
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