Thermo Scientific Pierce Rapid Mouse Antibody Isotyping Kits are single-use, cassette-based kits for 5-minute lateral-flow assay determination of mouse monoclonal antibody class and subclass identity.
These antibody isotyping kits use cartridges that provide a color-readout of the monoclonal antibody isotype within five minutes after pipetting a small amount of diluted culture supernatant or ascites fluid sample into the sample well. Two kit varieties are offered. One kit determines mouse IgG1, IgG2a, IgG2b, IgG3, IgA, IgM. The other kit determines those same subclasses as well as light chain identity (kappa vs. lambda light chains).
- Long shelf life – stable for at least one year at room temperature
- Single-step – add diluted antibody sample to the loading-well of cassette
- Fast – within 5 minutes a band appears that indicates the antibody isotype
- Sensitive – detect antibodies in tissue culture media or ascites fluid samples at any concentration greater than 10ng/mL
- Specific – assay does not cross-react with fetal bovine serum (FBS)
- Reliable – results are similar to the standard ELISA-based isotyping assays
Each Pierce Rapid Isotyping Kit assay is performed by simply adding a properly diluted tissue culture supernatant or mouse ascites sample to the well of the small cassette. Gold conjugates embedded in the cassette form specific class- and subclass-soluble complexes with the antibodies in the sample. These complexes travel the length of the membrane and are resolved on the anti-isotype and class-specific antibody-impregnated membrane. Results are displayed as a red band indicating the antibody isotype or subclass.
Determining the class and subclass of a monoclonal antibody is useful in planning the best immunoglobulin purification method. For example, mouse IgA and IgM are best purified by size (i.e., gel exclusion) or using immunoaffinity separation columns. Mouse IgG2a and IgG2b are purified with immobilized Protein A at pH 7-8, while Mouse IgG1 binds best to Protein A at pH 8-9. Immunoglobulin that contains kappa light chains can be purified using immobilized Protein L.
Pierce Rapid Isotyping Kits are compatible with both tissue culture supernatant and mouse ascites fluid. They are more sensitivity than conventional latex-bead based dipstick assays and much faster than ELISA-based isotyping assays. Kits are available for isotyping mouse IgG1, IgG2a, IgG2b, IgG3, IgA, IgM and kappa and lambda light chains and require only 0.5µL ascites fluid, 5 to 55µL of cell culture supernatant or 1.5ng of purified antibody.
Review of antibody screening, isotyping and quantitation methods
Rapid ELISA Antibody Isotyping Kit – for 1-hour isotyping of multiple samples
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