DiO/DPAmembranepotentialdetectionkitcomprisesavialofthegreenfluorescentmembranedyeDiOC16(3)dissolvedinDMSO(1mLat2mM)andavialoftheanionicmembranequencherdyeDPA(dipicrylamine)dissolvedinDMSO(1mLat20mM).TheDiO/DPAsystemdetectscytoplasmicmembranepotentialchangesusingtheprincipleoffluorescenceresonanceenergytransfer(FRET),whereDiOisa“stationary”FRETdonorwhileDPAactsasamobileFRETacceptor.ThemembranelocationofDPAisafunctionofthepolarityandmagnitudeofmembranepotential,thusresultinginamembranepotential-dependentFRETefficiency.TheDiO/DPAsystemhasbeenreportedtoproduceafluorescencesignalchangeof>56%inHEK-293cellsand>25%inneuronalculturesandbrainslicesper100mVmembranepotentialchange(Bradley,etal.J.Neurosci.29(29),9197(2009)).
TheDiOC16(3)inthiskitisanimprovedversionovertheDiOC16(3)fromothersuppliers;ourDiOC16(3)comprisesgluconateasthecounterion,whichmakesthedyemorewatersoluble,thusfacilitatingcellstaining.DPAistrADItionallyahazardousmaterialduetoitsexplosivenature.Toimproveitssafetyduringshippingandhandling,wehavemadeitavailableasadilutedsolutionorasasolidinasmallerandmoreconvenientpackagingunit(Seecatalogno.60037).
References
1.JNeurosci29(29),9197(2009).
