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Monitoring Receptor-Mediated Changes in [Ca2+]i Using Single-Wavelength and Dual-Wavelength Indicators on the FlexStation Workstation

aw, C., Lydford, S.J. & Henderson, D.J. (1998). Characterisation of the P2receptor(s) on ECV304 endothelial cells using FLIPR. Naunyn-Schmeidebergs Arch.Pharmacol. 358, R126.

8. Zambon, A.C., Conklin, B.R., Brunton, L.L., Insel, P.A. & Ferguson, A.T. (2001). Application Note #1. Gq Protein Coupled Receptor Assays Using FlexStation. (http://www.moleculardevices.com/pdfs/flexstation_1.pdf).

9. Zhang, J-H., Thomas, T.D.Y. and Oldenburg, K. R. (1999). A simple statistical parameter for use in evaluation and validation of high throughput screening assays. J. Biomol. Screening, 4, 6773.


APPENDIX
Materials
Cells : CHO cells (M1-WT3) - adherent Chinese Hamster Ovary cells stably expressing the rM1-muscarinic receptor (ECACC Ref. #: 940822109)

Reagents: FlexStation Calcium Assay Kit (Molecular Devices, cat # R8041), ATP (Adenosine 5'-triphosphate disodium salt, Sigma, cat # A2383), probenecid (Sigma, cat # P8761), DMSO, low water content (Sigma, cat # D2650), Fluo-3 AM ester (Molecular Probes, cat # F1242), Fura-2 AM ester (Molecular Probes, cat # F1221) and 20% pluronic acid solution (Molecular Probes, cat # P3000)

Plasticware : 75cm2 culture flasks (Corning cat # 430641), 96-well black walled, clear bottomed plates (Costar, cat # 3603), 96-well clear, U-bottom polypropylene plates (Greiner Bio-One, cat # 650101), Pipette tips for FlexStation - 200 L non-sterile, polypropylene tips (Molecular Devices, black cat # 9000-0622 or clear cat # 9000-0623)

Media and buffers : Hams-F12 medium (Invitrogen Life Technologies, cat # 31765-027), Hank's Balanced Salt Solution (HBSS) (10X) (Invitrogen Life Technologies, cat # 14065-049), HEPES 1M buffer solution (Invitrogen Life Technologies, ca
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