Phosph-Free Block ELISA Blocking Buffer

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.

Phosph-Free Block ELISA Blocking Buffer is a novel non-protein blocking formulation designed to eliminate interference and nonspecific background noise associated with antibody-coated ELISA formats and sandwich ELISAs., , This block buffer is formulated for ELISAs using alkaline phosphatase detection or for assays with ultra-sensitivity requirements. Because this ELISA buffer contains inert non-reactive blockers, it reduces the amount of nonspecific binding of enzyme labeled conjugates to blocked plate surfaces. Use of this blocking formulation minimizes backgrounds without the use of conventional cross-reactive protein additives., , Phosph-Free Block works on all types of polystyrene plates except Immulon® II microplates. When blocking with this buffer, ICT recommends Corning® 96-Well EIA/RIA StripWell™ microplates (ICT catalog #25).