Neptune Block Blocking Buffer is a non-mammalian blocking buffer designed primarily for antigen-down ELISA formats as well as sandwich assays with high background problems. Neptune Block is particularly useful when working with human and other mammalian serum samples as it works to reduce interactions between sample and blocking molecules.
Neptune Block ELISA Blocking Buffer is a mixture of small molecular proteins and molecular stabilizers capable of blocking the nonspecific binding sites of the adsorbed proteins, as well as the unoccupied regions of the plate that are not accessible to larger mammalian blocking reagents. When used in antigen-down ELISA formats, this product’s low molecular-weight proteins reduce the possibility of false-positives generated from endogenous antibodies in the sample reacting with plate-blocking proteins.
This buffer contains an antimicrobial agent for room temperature blocking. Bulk volumes and custom packaging are available upon request.
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.
• pH: 7.4 at 1X
• Contains: Non-Mammalian Proteins
• Supplied At: 1X
• Storage: 2-8°C
Abadir P, Jain A, Powell L, Xue QL, Tian J, Hamilton RG, Bennett D, Finucane T, Walston JD, Fedarko NS. Discovery and Validation of Agnostic Angiotensin Receptor Autoantibodies as Biomarkers of Adverse Outcomes. Circulation. Abstract.
“Wells were blocked with Neptune blocking reagent (ImmunoChemistry Technologies, Bloomington, MN, USA).”