Bpsbioscience/GAL4 Reporter Kit (Glucocorticoid Receptor Pathway)/60522/500 reactions188bio精品生物—专注于实验室精品爆款的电商平台

Background:

Theglucocorticoidsignalingpathwayplaysanimportantroleindevelopment,fluidhomeostasis,cognition,immuneresponseandmetabolism.Glucocorticoidsareaclassofsteroidhormonesthatbindtotheglucocorticoidreceptor,causingittotranslocatetothenucleus.Upontranslocation,thereceptorcanregulatethetranscriptionofalargenumberofgenes,includingthosethatregulateglucosemetabolismandinflammatoryresponses.

Description:

TheGlucocorticoidReceptorPathwayReporterKitisdesignedformonitoringtheactivityoftheglucocorticoidsignalingpathwayinculturedcells.Thekitcontainsatransfection-readyexpressionvectorfortheglucocorticoidreceptorligandbindingdomainthatisfusedtotheDNAbindingdomain(DBD)ofGAL4(GAL4DBD-GR).ThisfusionconstructactivatesfireflyluciferaseexpressionunderthecontrolofamultimerizedGAL4upstreamactivationsequence(UAS).Thisallowsforspecificdetectionofglucocorticoid-inducedactivationoftheglucocorticoidreceptorwithouttheneedforindividualtranscriptionaltargetsandwithlowcrossreactivityforothernuclearreceptorpathways.TheGAL4/UASreporterispremixedwithconstitutivelyexpressingRenilla(seapansy)luciferasevector,whichservesasaninternalpositivecontrolfortransfectionefficiency.

Thekitalsoincludesanon-induciblefireflyluciferasevectorpremixedwithconstitutivelyexpressingRenillaluciferasevectorasanegativecontrol.Thenon-inducibleluciferasevectorcontainsafireflyluciferasegeneunderthecontrolofaminimalpromoter,butwithoutanyadditionalresponseelements.Thenegativecontroliscriticalfordeterminingpathway-specificeffectsandbackgroundluciferaseactivity.

Synonym(s):

GlucocorticoidReceptor,GR,Nuclearreceptorsubfamily3groupCmember1,NR3C1,GAL4,GAL4/UAS

Format:

COMPONENTS:

Instructionsforuse:

Seeassaykitdatasheetfordetailedprotocol.

Storage/Stability:

>1yearfromdateofreceiptwhenstoredat-20°C

Application(s):

•Monitorglucocorticoidsignalingpathwayactivity.•Screenactivatorsorinhibitorsoftheglucocorticoidsignalingpathway.

Reference(s):

PaguioA,etal.(2010)ImprovedDual-LuciferaseReporterAssaysforNuclearReceptors.CurrChemGenomics.4:43-49.

Notes:

MaterialsRequiredbutNotSupplied
•Mammaliancelllineandappropriatecellculturemedium
•96-welltissuecultureplateor96-welltissueculturetreatedwhiteclear-bottomassayplate(Corning#3610)
•Transfectionreagentformammaliancellline.[WeuseLipofectamine™2000(LifeTechnologies#11668027),however,othertransfectionreagentsshouldworkequallywell.]
•Opti-MEMIReducedSerumMedium(LifeTechnologies#31985-062)
•Dualluciferaseassaysystem:Dual-Glo®LuciferaseAssaySystem(Promega#E2920):Thissystemassayscellsdirectlyingrowthmedium.Itcanbeusedwithanyluminometer.Automatedinjectorsarenotrequired.ORDual-Luciferase®ReporterAssaySystem(Promega#E1910):Thissystemrequiresacelllysisstep.Itisidealforluminometerswithautomatedinjectors.
•Luminometer

Thesevectorsarereadyfortransienttransfection.TheyareNOTSUITABLEfortransformationandamplificationinbacteria.

Warning(s):

Avoidfreeze/thawcycles.

ScientificCategory:

Cell-BasedAssayKit

ProductType:

AssayKit

Datashownislot-specific.Contactusforspecificinformationonotherlots.

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Bpsbioscience/GAL4 Reporter Kit (Glucocorticoid Receptor Pathway)/60522/500 reactions