TheRARαReporterCellularAssayPackprovidesallthekeyreagentsrequiredtomonitortheactivityofretinoicacidreceptoralpha(RARα).ThepackcontainstheRARαReporter(Luc)-HEK293CellLine,aluciferasereportercelllinethatcontainsafireflyluciferasegeneunderthecontrolofretinoicacidresponseelementsstablyintegratedintoHEK293cellsalongwithfulllengthhumanRARα(GenBankAccessionNo.NM_000964).Thiscelllineisfunctionallyvalidatedfortheresponsetothestimulationofall-transretinoicacid,andtheexpressionofRARαisconfirmedbyWesternblotting.
Additionally,thepackincludescellculturemedium(ThawMedium6)thathasbeenoptimizedforusewithHEK293cells.ThawMedium6includes10%fetalbovineserumand1%Pen/Strep.Finally,thepackprovidestheONE-Step™LuciferaseDetectionSystem.Thisreagentprovideshighlysensitive,stabledetectionoffirefly(Photinuspyralis)luciferaseactivity.TheONE-Stepluciferasereagentcanbeuseddirectlyincellsingrowthmedium,andcanbedetectedwithanyluminometer;automatedinjectorsarenotrequired.
•All-TransRetinoicAcid(ATRA)(Sigma-Aldrich,#R2625):stocksolutioninDMSO
•96-welltissueculturetreatedwhiteclear-bottomassayplate(Corning,#3610)
•Luminometer
ThawMedium6(BPSCat.#60183):DMEMmedium(Hyclone,#SH30243.01)supplementedwith10%FBS(Lifetechnologies,#26140-079),1%Penicillin/Streptomycin(Hyclone,SV30010.01).
CompleteGrowthMedium:ThawMedium6and400µg/mlofGeneticin(G418;Invitrogen,#11811031),1µg/mlofPuromycin(Hyclone,#SV30075.01),and100µg/mlHygromycin(Hyclone,#SV30070.01).
•ScreenagoNISTsorantagonistsofRARα.
1.Petkovich,M,etal.Nature(1987)330(6147):444-450.
2.Allenby,G,etal.Proc.Natl.Acad.Sci.USA(1993)90(1):30-34.
LicenseDisclosure
Purchaseofthiscelllinegrantsyouwitha10-yearlicensetousethiscelllineinyourimmediatelaboratory,forresearchuseonly.Thislicensedoesnotpermityoutoshare,distribute,sell,sublicense,orotherwisemakethecelllineavailableforusetootherlaboratories,departments,researchinstitutions,hospitals,universities,orbiotechcompanies.Thelicensedoesnotpermituseofthiscelllineinhumansorfortherapeuticordruguse.Thelicensedoesnotpermitmodificationofthecelllineinanyway.InappropriateuseordistributionofthiscelllinewillresultinrevocationofthelicenseandresultinanimmediateceaseofsalesanddistributionofBPSproductstoyourlaboratory.BPSdoesnotwarrantthesuitabilityofthecelllineforanyparticularuse,anddoesnotacceptanyliabilityinconnectionwiththehandlingoruseofthecellline.Modificationsofthiscellline,transfertoanotherfacility,orcommercialuseofthecellsmayrequireaseparatelicenseandadditionalfees;contactsales@bpSBIoscience.comfordetails.PublicationsusingthiscelllineshouldreferenceBPSBioscience,Inc.,SanDiego.
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