ELISA Assay Buffer (5X)

Each CytoSets contains capture (coating) antibody, biotinlyated detection antibody, standard and Streptavidin-HRP. Other reagents required are listed in the CytoSets information sheet included with the kit and can be purshased from us separately (Antibody Pair Buffer kit CNB0011, 5x Assay Buffer DS98200, etc.). The information sheet also provides a specific procedure and illustrates an example standard curve which can be obtained when the specific procedure is followed. A general procedure is summarized here:

1) Coat the microplate with diluted capture (coating) antibody overnight at 2-8 degrees C; Wash the plate
2) Incubate standards or samples with the coated microplate; Wash the plate
3) Incubate diluted biotinlyated detection antibody with the plate; Wash the plate
4) Incubate Streptavidin-HRP with the plate for 15-45 minutes; Wash the plate
5) Incubate the plate with TMB substrate for 10-60 minutes and stop the reaction with Stop solution
6) Read microplate at 450 nm.

Investigators are advised to determine optimal buffer formulations, concentrations and incubation times for individual applications.

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