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Journal of Biomolecular Screening
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Monitoring Inositol-Specific Phospholipase C Activity Using a Phospholipid FlashPlate®

Thomas R. Mullinax

NEN Life Science Products, Boston, MA

Georgette Henrich

NEN Life Science Products, Boston, MA

Patricia Kasila

NEN Life Science Products, Boston, MA

David G. Ahern

NEN Life Science Products, Boston, MA

Elizabeth A Wenske

NEN Life Science Products, Boston, MA

Cuifen Hou

The R.W. Johnson Pharmaceutical Research Institute, Raritan, NJ

Dennis Argentieri

The R.W. Johnson Pharmaceutical Research Institute, Raritan, NJ

Michael E. Bembenek

NEN Life Science Products, Boston, MA

Inositol-specific phospholipase Cs(PLCs) are a group of enzymes involved in the signal transduction pathway of many plasma membrane receptor mediated events. We developed a modified solid surface to capture [3H] PIP2 onto the Basic FlashPlate® in order to monitor PLC activity. Our results clearly demonstrate the utility of [3H] PIP2-Coated Phospholipid FlashPlate® microtiter plates for assessing PLC activity for HTS of receptor-coupled functional assays. The results show that PLC activity can be measured easily from a variety of sources including purified recombinant enzyme preparations, crude HL60 cell lysates and permeabilized A431 human carcinoma cells. Moreover, this format provides a surface comparable to that used for classical solution based radiolabeled mixed phospholipid micelle studies and illustrates the feasibility of this assay for measuring PLC activation in a variety of different drug screening assays.

Journal of Biomolecular Screening, Vol. 4, No. 3, 151-155 (1999)
DOI: 10.1177/108705719900400309
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