Alternative 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.

Alternative Block is a protein-free, detergent-free ELISA blocking buffer. This unique blocking buffer contains a heterogeneous mixture of proprietary blockers and synthetic stabilizers that block the uncoated regions of the microtiter plate without the use of conventional cross-reactive protein additives or detergents. This buffer is best for most immunoassays, including antibody-sandwich ELISA, competitive assays, and peptide and protein antigen-down ELISA., , Alternative Block’s all-synthetic heterogenous mixture of blocking agents minimizes non-specific binding interactions during the immunoassay process to reduce background noise. Alternative Block provides a microhydrated environment to stabilize the coated protein during long-term storage for improved retention of antigen epitope and antibody binding activity. When preparing batches for storage, an antimicrobial agent allows for stable room temperature blocking and long-term refrigerated storage of dried plates.