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This standard operating procedure describes the methods for determining the optimal concentration of a relevant capture antibody or soluable antigen and lot of specific enzyme linked secondary antibody. This document also contains methods for determining the suitability for use of a lot of substrate buffer and PNPP substrate, as measured against lots of known performance. This procedure is applicable for use in quantitation of 12.8 antibody in solutions, and can be used in the evaluation of reagents for most ELISA procedures.