Secreted alkaline phosphatase (SEAP) is a truncated form of the human pla-cental alkaline phosphatase lacking a membrane anchoring domain. The enzyme is secreted from the cell, allowing analysis using a small amount of the cell culture medium. This offers the major advantage that the cells remain intact for further analyses. Also, because SEAP is extremely heat stable, endogenous phosphatase activity can be eliminated by a simple heat treatment of the sample at 65°C. The chemiluminescent assay for SEAP is very sensitive, dephosphorylation of the chemiluminescent phosphatase substrate (CSPD) results in a glow that is measured in a luminometer and remains constant for up to 60 min. We use a commercial assay kit to measure SEAP activity, the Roche SEAP Reporter Gene Assay.
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