Trouble-free determination of mouse antibody class and subclass identity for single or multiple samples.

The Thermo Scientific Pierce Rapid ELISA Mouse mAb Isotyping Kit enables easy identification of mouse immunoglobulin class, subclass and light chain. This fast assay uses ELISA strip-plates 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. 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 450 nm.

Highlights

• Fast – determine antibody isotype in ~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 >3 ng/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 (450 nm)
• Complete and easy to use – includes precoated plates, detecting antibody, buffers, and TMB substrate and stop solutions

Procedure for the Thermo Scientific Pierce Rapid ELISA 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 30-60 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 Pierce 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 50 µl 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.

Related Products and Resources:

Rapid ELISA Mouse Antibody Isotyping Kit FAQ
Rapid Antibody Isotyping Kits (single-use "cassettes" for isotyping in 5 minutes!)
Easy-Titer Antibody Assay Kits (to determine antibody concentration)
Protein A, G, A/G and L Antibody Purification Kits
Melon Gel Monoclonal Antibody Purification Kit