ICT’s Neptune Block Blocking Buffer is non-mammalian and designed primarily for antigen-down ELISA formats as well as sandwich assays with high background problems. This buffer is particularly useful when working with human and other mammalian serum samples as it works to reduce interactions between sample and blocking molecules.
- Coat antibody or antigen onto the ELISA plate using ICT’s Antibody Coating Buffer or Antigen Coating Buffer.
- Incubate 8–24 hours at room temperature (RT).
- Aspirate the coating solution.
- Wash each well twice with ICT’s ELISA Wash Buffer.
- 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.
- Incubate 8–24 hours at RT. For best blocking, incubate overnight at RT.
- Aspirate the blocking buffer; do not wash.
- 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.
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