Pro-Detect™ Rapid Antibody Isotyping Assay Kit, mouse (Thermo Scientific™)

The Thermo Scientific Pro-Detect Rapid Antibody Isotyping Assay Kit - Mouse is a single-use, dipstick lateral flow kit for quick, easy determination of mouse monoclonal antibody class and subclass identity. This antibody isotyping assay uses membrane-based strips coated with dye-bound conjugated antibodies that provide a visual, color readout of the monoclonal antibody isotype within ten minutes. The kit determines mouse IgG1, IgG2a, IgG2b, IgG3, IgA, IgM, as well as light-chain identity (kappa vs. lambda light chains).

Features of the Pro-Detect Rapid Antibody Isotyping Assay Kit:
• Long shelf life—stable for at least one year at 4°C
• Simple workflow—dilute antibody sample using provided diluent and dip in the lateral flow strip
• Fast—within 5–10 minutes a band appears that indicates the antibody isotype
• Sensitive—detect antibodies in tissue culture media or ascites fluid samples down to low ng/mL concentrations
• Specific—assay does not cross-react with fetal bovine serum (FBS)
• Reliable—results are similar to standard ELISA-based isotyping assays

Each Pro-Detect Rapid Antibody Isotyping assay is performed by simply adding a properly diluted tissue culture supernatant or mouse ascites sample to a test tube or microplate well and dipping in a lateral flow strip. Gold conjugates embedded in the strip membrane 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 regions of the membrane. Results are displayed as a red band above the printed class or sub-class description, indicating the antibody isotype present.

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.

The Pro-Detect Rapid Antibody Isotyping Assay Kit is compatible with both tissue culture supernatant and mouse ascites fluid, and is more sensitive than conventional latex bead-based dipstick assays and much faster than ELISA-based isotyping assays. The kit requires only 0.5 μL ascites fluid, 2–22 μL of cell culture supernatant or 1.5 ng of purified antibody.