Abstract
Scintillation proximity assay (SPA) is a bead-based homogeneous assay technology that removes the need for a filtration step to separate bound from free ligand in a receptor binding assay. SPA allows the rapid and sensitive assay of a wide range of molecular interactions in a homogeneous system and is routinely used for radioligand binding assays, particularly in drug screening applications where high throughput is required. Existing filter binding assays may be readily converted to SPA assays or assays may be directly developed in SPA format. This chapter describes the development of SPA radioligand binding assays detailing the choice of isotope, selection of SPA bead type, optimization of SPA bead and receptor ratio, optimization of assay buffer, selection of assay format, and assay validation including saturation binding, competition binding, and association/dissociation binding studies using SPA.
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 Tween is a trademark of ICI Americas, Inc.
References
Bosworth N, Towers P (1989) Scintillation proximity assay. Nature 341(6238):167–168
Anon, SPA bibliography. PerkinElmer. http://www.perkinelmer.com/spa
Anon, Determination of [35S]GTPγS binding by scintillation proximity assay (SPA), Proximity News 25. http://www.perkinelmer.com/spa
Berry JA, Burgess AJ, Towers P (1991) Scintillation proximity assay; competitive binding studies with [125I]endothelin-1 in human placenta and porcine lung. J Cardiovasc Pharmacol 17(7):S143–S145
Anon, Biotinylation techniques and their use in developing scintillation proximity assays, Proximity News 39. http://www.perkinelmer.com/spa
Anon, SPA cytokine receptor binding assays, Proximity News 37. http://www.perkinelmer.com/spa
Urban F Jr, Cavazos G, Dunbar J, Tan B, Escher P, Tafuri S, Wang M (2000) The important role of residue F268 in ligand binding by LXRβ. FEBS Lett 484:159–163
Anon, Selection of SPA bead type for receptor scintillation proximity assays, Proximity News 54. http://www.perkinelmer.com/spa
GraphPad Prism version 4.00 for Windows. GraphPad Software, San Diego, California. http://www.graphpad.com. Copyright © 1994–1999 by GraphPad Software
Anon, Kinetics of inhibition of receptor binding measured by scintillation proximity assay, Proximity News 14. http://www.perkinelmer.com/spa
Anon, Characterisation of d-myo-[3H]inositol 1,4,5 trisphosphate binding to purified inositol trisphosphate receptors by scintillation proximity assay, Proximity News 12. http://www.perkinelmer.com/spa
Patel S, Harris A, O’Beirne G et al (1995) Characterisation of inositol 1,4,5 trisphosphate binding by scintillation proximity assay. Br J Pharmacol 115(Proceedings Supplement):35P
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Up-to-date bibliography of SPA publications available at http://www.perkinlemer.com/citations.
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Berry, J., Price-Jones, M., Killian, B. (2012). Use of Scintillation Proximity Assay to Measure Radioligand Binding to Immobilized Receptors Without Separation of Bound from Free Ligand. In: Davenport, A. (eds) Receptor Binding Techniques. Methods in Molecular Biology, vol 897. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-61779-909-9_4
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DOI: https://doi.org/10.1007/978-1-61779-909-9_4
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