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Coating Buffers

A system of buffers for efficient and reliable coating of plates with antibodies or antigens.

BulletCoating Buffers -

The first step in making a reliable ELISA is the proper coating of the antibody or antigen onto the plate. HDB offers 2  plate coating buffers for use with either antibodies or antigens. Both coating buffers stabilize coated proteins by maintaining their tertiary three-dimensional structure, allowing for greater binding reactivity with the detection molecule, thereby enhancing the specific signal.

 

For antibody-sandwich ELISAs, use the 5x Universal Antibody Plate Coating Buffer, a unique buffer designed for coating antibodies onto polystyrene ELISA plates. This buffer can also be used for antigen-down ELISAs as well. However, for most antigen-down ELISAs, we recommend the 5x Antigen Coating Buffer, which is optimized for antigen-down ELISAs.

 

Bullet5x Universal Antibody Coating Buffer -

The pH and ionic strength of this buffer have been specifically formulated to create an optimal coating environment which maximizes the adsorption efficiency of most antibodies and some protein antigens onto polystyrene plate surfaces.

 

This buffer also stabilizes coated antibodies by maintaining their tertiary three-dimensional structure during the adsorption process. By stabilizing the adsorbed antibody, antigenic regions are preserved, allowing for greater binding reactivity with the detection molecule, thereby enhancing the specific signal.

By generating a higher specific signal, a lower concentration of coating antibody may be needed when this buffer is used, thereby saving valuable reagents. There is a finite quantity of antibody that may be coated effectively onto microtiter plates.  Use of antibody concentrations in excess of the optimal range will result in surface saturation and clumping on the plate surface.  This produces lower specific signals, and higher nonspecific signals.

 

CATALOG # DESCRIPTION QTY PRICE
PURCHASE
REA-6245 Ab Coating Buffer 5X 25 ml
$ Call us
REA-644 Ab Coating Buffer 5X 100 ml
$ Call us
REA-645 Ab Coating Buffer 5X 500 ml
$ Call us
REA-646 Ab Coating Buffer 5X 1 liter
$ Call us
REA-658 Ab Coating Buffer 5X 10 liters
$ Call us
 
Bullet5x Antigen Coating Buffer -

This buffer is well suited to coat all antigens onto polystyrene plates and enhances coating adsorption by using an alkaline pH binding environment so that this buffer can be used for all antigen-down ELISAs. During the adsorption process, the Antigen Coating Buffer stabilizes and hydrates coated antigens without compromising their three-dimensional structure. This allows for greater accessibility to detector antibody that are present in sample solutions, thereby enhancing the specific signal. By generating a higher specific signal, a lower concentration of coat antigen may be needed when this buffer is used, thereby saving valuable reagents.

 

All antibody coating buffers contain an antimicrobial agent to retard bacterial growth during the plate coating process, allowing the procedure to be performed at room temperature. Depending on the activity of the coated antibody or antigen, plates may be stored at 2°-8°C (or even at room temperature for hardy proteins) for several months, or even years.


The coating buffers are supplied as a 5x concentrate to conserve valuable storage space. Simply add 4 parts deionized water to 1 part buffer, mix, and it’s ready to use. No pH adjustment is required. After dilution, the 1x buffer will suppress bacterial growth for up to 1 month at room temperature and 6 months at 2-8°C. The concentrated 5x buffer will suppress bacterial growth up to 18 months at 2-8°C. During refrigeration, some salts in the buffer may precipitate. Simply warm the bottle in a water bath (do not boil) and mix. The crystals should go into solution.

 

CATALOG # DESCRIPTION QTY PRICE
PURCHASE
REA-6246 Antigen Coating Buffer 5X 25 ml
$ Call us
REA-6247 Antigen Coating Buffer 5X 100 ml
$ Call us
REA-6248 Antigen Coating Buffer 5X 500 ml
$ Call us
REA-6249 Antigen Coating Buffer 5X 1 liter
$ Call us
REA-6250 Antigen Coating Buffer 5X 10 liters
$ Call us