Creative Diagnostics RapidGetTM ELISA Mouse mAb Isotyping Kit provides antigen-independent determination of mouse monoclonal antibody isotype using a microplate method that enables multiple samples to be tested in about one hour.
This fast assay uses ELISA strip-plates that are pre-coated in different wells with anti-mouse heavy-chain capture antibodies (anti-IgG1, IgG2a, IgG2b, IgG3, IgA and IgM) or anti-mouse light-chain capture antibody (kappa or lambda). This approach eliminates the need to purify and immobilize an antigen to determine immunoglobulin isotype (antibody subclass and light chain identity). A mouse monoclonal antibody sample applied to the wells can be isotyped within one hour. Results are evaluated qualitatively by visual inspection or quantitatively by measuring the absorbance at 450nm.
Protocol summary for the Creative Diagnostics RapidGetTMELISA Mouse mAb Isotyping Kit. The simple ELISA procedure involves only one probing-incubation step. Sample and detection antibody are added together to the wells of the precoated microplate. (Each well in an eight-well strip is coated with a different type-specific capture antibody.) After 30 to 60 minutes, TMB substrate is added to reveal the antibody isotype based on which wells in the strip produce color.
Pre-coated strip-well plates for the RapidGetTM Rapid ELISA Isotyping Kit clearly identify monoclonal antibodies. Wells A-H of each eight-well strip (microplate column) are pre-coated with eight different class- or subclass-specific capture antibodies. A sample is tested by adding 50µL of sample to all eight wells of a strip. For partial-plate assays with less than 12 samples, individual 8-well strips can be removed from the microplate frame for later use. In this example, the results indicate that the antibody tested in strip #1 is a mouse IgG1 with kappa light chains.