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dc.contributor.authorRossi, Ana
dc.contributor.authorTaylor, Colin
dc.date.accessioned2019-01-17T00:30:37Z
dc.date.available2019-01-17T00:30:37Z
dc.date.issued2020
dc.identifier.issn1064-3745
dc.identifier.urihttps://www.repository.cam.ac.uk/handle/1810/288097
dc.description.abstractFluorescence polarization (FP) can be used to measure binding of a small fluorescent ligand to a larger protein because the ligand rotates more rapidly in its free form than when bound. When excited with plane polarized light, the free fluorescent ligand emits depolarized light, which can be quantified. Upon binding, its rotation is reduced and more of the emitted light remains polarized. This allows FP to be used as a nondestructive assay of ligand binding. Here we describe a fast, high-throughput FP assay to quantify the binding of fluorescently labeled inositol 1,4,5-trisphosphate (IP3) to N-terminal fragments of the IP3 receptor. The assay is fast (1-6 h), it avoids use of radioactive materials and when measurements are performed at different temperatures, it can resolve Gibbs free energy (ΔG°), enthalpy (ΔH°), and entropy (ΔS°) changes of ligand binding.
dc.format.mediumPrint
dc.languageeng
dc.publisherSpringer US
dc.subjectInositol 1,4,5-Trisphosphate
dc.subjectLigands
dc.subjectFluorescence Polarization
dc.subjectThermodynamics
dc.subjectEntropy
dc.subjectInositol 1,4,5-Trisphosphate Receptors
dc.titleAnalyses of Ligand Binding to IP3 Receptors Using Fluorescence Polarization.
dc.typeArticle
prism.endingPage120
prism.publicationDate2020
prism.publicationNameMethods Mol Biol
prism.startingPage107
prism.volume2091
dc.identifier.doi10.17863/CAM.35412
dcterms.dateAccepted2018-11-07
rioxxterms.versionofrecord10.1007/978-1-0716-0167-9_9
rioxxterms.versionAM
rioxxterms.licenseref.urihttp://www.rioxx.net/licenses/all-rights-reserved
rioxxterms.licenseref.startdate2020-01
dc.contributor.orcidTaylor, Colin [0000-0001-7771-1044]
dc.identifier.eissn1940-6029
rioxxterms.typeJournal Article/Review
pubs.funder-project-idBiotechnology and Biological Sciences Research Council (BB/P005330/1)
pubs.funder-project-idWellcome Trust (101844/Z/13/Z)
cam.issuedOnline2019-11-27
cam.orpheus.successThu Jan 30 10:53:04 GMT 2020 - Embargo updated
rioxxterms.freetoread.startdate2021-01-31


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