Antigen Coating Buffer, 5X

1. Coat antibody or antigen onto the ELISA plate using ICT’s Antibody Coating Buffer or Antigen Coating Buffer., 2. Incubate 8–24 hours at room temperature (RT)., 3. Aspirate the coating solution., 4. Wash each Well twice with ICT’s ELISA Wash Buffer., 5. Block the uncoated regions of the ELISA plate by pipetting 300–400 µL of blocking buffer into each Well. Always use an equal or greater volume of blocking buffer than was used for the coating buffer solution., 6. Incubate 8–24 hours at RT. For best blocking, incubate overnight at RT., 7. Aspirate the blocking buffer; do not wash., 8. Run the assay immediately, or dry the plate for long-term storage and seal in a foil storage bag with a desiccant pack. Store dried and packaged plates at 2-8°C.

Antigen Coating Buffer, 5X maximizes the adsorption of antigens onto polystyrene plates for use in an indirect ELISA. By stabilizing the tertiary structure of the antigen, Antigen Coating Buffer helps the antigen retain its binding reactivity with the target antibody in the sample. This product is best for coating proteins ranging from 6 kDa to > 200 kDa onto polystyrene plates. Stabilizing the coated antigen allows for better analyte detection and enhances the specific signal of the assay. By generating a higher positive signal, a lower concentration of coating antigen may be used, saving valuable reagents. Under proper storage conditions, coated plates may last for years, enabling the manufacture of large batches for inclusion in diagnostic test kits. This also proves useful in non-manufacturing settings, where large batches of plates may be prepared at once to be stored for future experiments.