RapidGet™ ELISA Mouse Antibody Isotyping Kit

Creative Diagnostics RapidGet^TM 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.

Highlights:

• Fast – determine antibody isotype in approx. 1 hour; considerably faster than traditional ELISA kits
• Convenient – eight-well strip format allows partial use of the plate; use one strip (column) for each sample (12 samples per plate)
• Specific – characterize antibodies for six different subclasses and two different light-chain types
• Sensitive – accurate characterization with samples containing at least 3ng/mL of test antibody
• Flexible – kit is compatible with hybridoma cell culture supernatant, ascites fluid or purified antibodies
• Cost effective – cost per assay is less than with single-use assay strips or cassettes
• No special equipment needed – assess results visually or measure quantitatively using an ordinary ELISA plate reader (450nm)
• Complete and easy to use – includes precoated plates, detecting antibody, buffers, and TMB substrate and stop solutions

Protocol summary for the Creative Diagnostics RapidGet^TMELISA 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 RapidGet^TM 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.