Our Coating and Wash Buffers play an important role in optimizing your ELISA assay performance.
Coating Buffers: what are they and how do they work?
The two most common types of ELISA assays are the antigen-down ELISA (in which the antigen of interest is immobilized onto the ELISA plate) and the antibody-sandwich ELISA (in which a capture antibody is immobilized onto the ELISA plate). For more details, see our Guide to Developing Optimized ELISAs.
To ensure the best ELISA performance, it is crucial that adsorption of the antigen or capture antibody onto the surface of the ELISA plate is optimized.
Coating Buffers perform two functions. Firstly, they enhance adsorption of the antigen or antibody to polystyrene ELISA plates. Secondly, they help to stabilize the 3-D structure of the immobilized antigen or antibody and thus maintain the functional integrity of the antigen epitope(s) and the antibody binding sites. In doing so, Coating Buffers enhance the specificity of the assay and help extend the shelf-life of coated plates by preventing degradation / denaturation of the immobilized proteins during plate storage.
By both maximizing antigen / antibody adsorption and stabilizing the immobilized protein, Coating Buffers can also reduce the amount of valuable, and often scarce, antigen or antibody that is required for each plate – an important consideration when preparing large quantities of plates.
Antibody Coating Buffer and Antigen Coating Buffer: what is the difference?
Both buffers are designed to perform the same function - to optimize the coating of polysterene ELISA plates.
- Antigen-Down ELISA: As the names suggests, our Antigen Coating Buffer is specially formulated to work with peptide and protein antigens, and helps retain the functional integrity of epitopes in the immobilized antigen. This is the ideal buffer for coating plates to be used in the antigen-down format. We recommend using the Immulon™ II HB 96-Well Microtiter EIA Plate for this ELISA format.
- Antibody-Sandwich ELISA: Similarly, our Antibody Coating Buffer is specially formulated to work with antibodies, and is formulated to help retain the functional integrity of antigen-binding sites on the immobilized antibody. This is the ideal buffer for coating plates to be used in antibody-sandwich format. We recommend using the Costar®96-Well EIA/RIA Stripwell Plate for this ELISA format.
Why is the wash buffer important?
When building, developing and running an ELISA assay, it is vital not to overlook the importance of correctly washing the plate between each reagent addition step. Thorough washing is important to remove interfering substances, reduce background, and maximize the specific signal. Therefore, it is important to use a high-quality wash buffer to ensure optimal washing of the plate. A poorly washed plate will result in a poorly performing ELISA.
Why use ICT's Wash Buffer?
ImmunoChemistry Technologies has developed ELISA Wash Buffer, 10X as a universal ELISA wash buffer that can be used (diluted to 1X with diH2O) with both antigen-down and antibody-sandwich ELISA formats.
It contains a specially formulated combination of pH stabilizers, salts, and detergents that effectively remove unbound proteins and assay components from the ELISA plate whilst maintaining specific antigen-antibody interactions. In this way it reduces background noise and maximizes the specific signal. It is a convenient and effective reagent for all your ELISA washing steps.
Learn more about our coating buffers
Learn more about our wash buffer
View our ELISA and Assay Reagents Catalog
View our ELISA and Assay Reagents Advantages Brochure
View our Guide to Developing Optimized ELISAs
Immulon is a trademark of ThermoLabsystems. Costar is a trademark of Corning, Inc.