The Thermo Scientific Pro-Detect Rapid Antibody Isotyping Assay Kit - Rat is a single-use, dipstick lateral flow kit for quick, easy determination of rat 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 rat IgG1, IgG2a, IgG2b , and IgG2c.

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 subclass of a rat antibody is important when selecting the best purification method. IgG2a, IgG2b, and IgG2c can be purified with Protein A columns at a pH of 7–8. IgG1 isotypes bind best to Protein A at a slightly higher pH of 8–9.

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.