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Monitor Purified Phosphatase Activity with a Homogeneous,
Non-Radioactive High-Throughput Fluorogenic Assay
Said Goueli, Ph.D., Kevin Kupcho, B.S., and Kevin Hsiao, M.S.,
Promega Corporation
We have developed a homogeneous, non-radioactive rapid assay system for
monitoring the activity of purified serine/threonine protein phosphatases (S/T
PPases), the ProFluor™ Ser/Thr PPase Assay. The robust assay performance is
manifested by its high Z´-factor values and its excellent dynamic range. The
protocol can be completed in as little as two hours, requires only two reagent
additions, and the signal is stable for hours. This assay is scalable from
single tubes to 384-well plates and is adaptable to automated platforms. We have
validated this assay with known selective inhibitors of protein phosphatases.
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