How to Reduce Non-Specific Binding to Containers

Coat the plate wells with the antigen. Use an antigen dilution. Pure solutions of antigen are not required since only about 3 percent of the protein in the sample is the target protein. When diluting, ensure that the samples contain the antigen at a concentration that is within the detection range of the antibody. Pipette the antigen dilution solution down the plate wells. Some plates may be pre-coated with antigen. Read the instructions supplied with the plate wells.

Cover the plates with an adhesive plastic and incubate overnight. Wash the plates with buffer solutions, such as Tris-buffered saline or phosphate-buffered saline. To wash, rinse the plates using a squeeze bottle filled with the buffer solution over a laboratory sink. Pat the plates with a paper towel to remove any remaining drops of buffer solution.

Reduce the non-specific binding of proteins and molecules using a blocking buffer or reagent, such as a detergent blocker, dry milk or bovine serum. The type of blocking reagent used will depend on the assay. Add the blocking reagent to the wells using a pipette. Cover the plate again with adhesive plastic and incubate overnight.

Re-wash the plate wells again using a buffer solution. These steps will decrease the amount of non-specific binding that can occur to the plate wells before adding the necessary antibody solution for the procedure. In addition, the blocking reagents can help reduce background colors in the results of the assays.