Recommended concentrations for use are approximate values. A dose dependent response assay should be performed to determine the optimal concentration for use in specific applications.ELISA and Western ELISA require 10-100ng protein depending on the nature and affinity of the detection reagent. Leptospira converted human serum polyclonal antibodies yield titers of 1:1000 or greater at 100ng of immobilized protein under standard ELISA conditions.