Preparation of microwell plates involve binding the antigen or antibody to the plate, blocking nonspecific binding sites, then coating the plate with a stabilizing reagent to allow for long term storage. ZeptoBind is specifically formulated to aid in the binding of antigens or antibodies to microwell plates. The reagent is supplied in a liquid, ready-to-use format.
The concentration of antibody or antigen will greatly affect the quality of the assay being developed. We recommend titrating the amount of antigen or antibody to be used for coating the wells. For most antibodies or antigens, the optimal coating concentration is in the range of 1 to 10 ug/mL when using ZeptoBind. If the optimal concentration is unknown and it is not practical to perform a titration to determine this concentration, we recommend using 10 ug/mL, which works well for most applications. A complete discussion of optimization procedures can be found in Harlow and Lane, “Antibodies, A Laboratory Manual”, Cold Spring Harbor Laboratory, 1988.
Dilute antigen or antibody in ZeptoBind. Add 100 uL of the suspension to each microwell and incubate overnight at 4°C. Aspirate the remaining solution. It is recommended that after binding the antibody or antigen to the plate, that a blocking reagent such as ZeptoBlock (Catalog # 0801185) and a stabilizing reagent such as ZeptoCoat (Catalog # 0801179) are applied. After binding the antigen or antibody to the plate, add 200 uL of ZeptoBlock to each well. Incubate for 2 hours at room temperature. Aspirate remaining solution then add 100uL of ZeptoCoat to each well. Incubate for 1 hour at room temperature. Aspirate the remaining solution and allow the plate to dry at room temperature for a minimum of 4 hours or overnight. Seal in a bag with a desiccant and store at 2-8 °C.
STORAGE AND STABILITY
Store at 2-8 °C. ZeptoBind contains no preservatives. Use aseptic technique when removing contents to avoid possible contamination. Best if used before expiration date (see product label).