TheDurhampHScreenMD1-101
TheDurhampHScreenisaproteinstABIlityscreendesignedbyEhmkePohl,MortenGrøftehaugeandEmilyCardewatDurhamUniversitytobeusedwithaproteinstabilityassay,suchasthethermalshiftassay(alsoknownasThermoFluor®).ThescreenidentifieshowbufferchemistryandpHaltersthethermaldenaturationandthereforestabilityofaprotein.Inaddition,buffermoleculescanactaslowaffinityligandsandbindtospecificpocketsonaprotein,thiscanbeanundesirableconfoundingfactorinfragment-baseddrugdesignprojects.Therefore,aproteinstabilityscreenfortheoptimumbufferandpHforyoursampleisavitalfirststepinanystructuredeterminationproject.
TheDurhampHScreenincludes28differentbufferscoveringabroadpHrange(4-11),andseparatestheeffectsofbufferchemistryandpH.Werecommendit"susetogetherwiththeDurhamSaltandRUBICsscreenstothoroughlyunderstandtheeffectofproteinenvironmentonstability.
TheDurhamScreensaredesignedtoworkwithNAMI*thefreetodownloadsoftwareforeasyinterpretationofthermalshiftassayresults.
Reference:Grøftehauge,MK,Hajizadeh,NR,Swann,MJandPohl,E.Protein-ligandinteractionsinvestigatedbythermalshiftassays(TSA)anddualpolarizationinterferometry(DPI).ActaCrystD71:36-44(2015).
TheDurhampHScreenispresentedas96x0.5mLconditionsinadeep-wellblock.Theyarepreservativefreeanditisrecommendedthattheyarestoredat4°C,howevertheycanbeusedatroomtemperatureforscreenset-up.
*NAMIcangeneratewaterfallplotsofthermalshiftdataatmultipleconcentrationsorplotsofmeltingtemperatureagainstconcentration.Thesedifferentiateasuddenincreaseinproteinstabilityindicatingaspecificinteractionandsteadyincreasesinstabilityduetothechangingproteinenvironment.Inaddition,itproducesausefulcolourgrADIentplatediagramtomakeidentifyingparticularlystabilisingordestabilisingconditionssimple.
ThermoFluorisaregisteredtrademarkofJohnsonandJohnson.