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Review

IP-3/IP-1 Assays

Kim E. Garbison  1 , Beverly A. Heinz, Mary E. Lajiness
Sarine Markossian  1 , Abigail Grossman  1 , Hannah Baskir  1 , Michelle Arkin  2 , Douglas Auld  3 , Christopher Austin  4 , Jonathan Baell  5 , Kyle Brimacombe  1 , Thomas D.Y. Chung  6 , Nathan P. Coussens  7 , Jayme L. Dahlin  8 , Viswanath Devanarayan  9 , Timothy L. Foley  10 , Marcie Glicksman  11 , Kirill Gorshkov  12 , Stefan Grotegut  13 , Matthew D. Hall  1 , Samuel Hoare  14 , James Inglese  1 , Philip W. Iversen  15 , Madhu Lal-Nag  16 , Zhuyin Li  12 , Jason R. Manro  13 , James McGee  17 , Allison Norvil  13 , Mackenzie Pearson  13 , Terry Riss  18 , Peter Saradjian  19 , G. Sitta Sittampalam  20 , Michael A. Tarselli  21 , O. Joseph Trask Jr.  22 , Jeffrey R. Weidner  23 , Mary Jo Wildey  24 , Kelli Wilson  1 , Menghang Xia  1 , Xin Xu  1
, editors.
In: Assay Guidance Manual [Internet]. Bethesda (MD): Eli Lilly & Company and the National Center for Advancing Translational Sciences; 2004.
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Affiliations
Review

IP-3/IP-1 Assays

Kim E. Garbison et al.

Excerpt

Activation of G-protein coupled receptors (GPCR) that couple to Gαθ and Gβγ and subsequent activation of phospholipase C –β (PLC-β) can be detected through the measurement of D-myo-inositol 1,4,5-triphosphate (IP3) in cells. This chapter describes technologies that can be used to develop robust assays for screening compounds, more specifically the use of Homogeneous Time-Resolved Fluorescence Assay (HTRF) for IP3 measurement. A sample preparation protocol, parameters for assay optimization and examples of data analysis are provided.

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