RecombinantJurkatTcellexpressingfireflyluciferasegeneunderthecontrolofNFATresponseelementswithconstitutiveexpressionofhumanPD-1(ProgrammedCellDeath1,PDCD1,SLEB2,CD279,GenBankAccession#NM_005018).
TCR-activatorPD-L1CHOcellline(BPSCat.#60536)
Assaymedium:ThawMedium3(BPSCat.#60186)
Anti-PD-1neutralizingantibody(BPSCat#71120)
Anti-PD-L1neutralizingantibody(BPSCat#71213)
96-welltissueculture-treatedwhiteclear-bottomassayplate
One-Stepluciferaseassaysystem(BPSCat#60690)orotherluciferasereagentsformeasuringfireflyluciferaseactivity
Luminometer
ThecelllinehasbeenscreenedusingthePCR-basedVenor®GeMMycoplasmaDetectionkit(Sigma-Aldrich)toconfirmtheabsenceofMycoplasmaspecies.
ThawMedium2(BPSCat.#60184):RPMI1640medium(LifeTechnologies#A10491-01)supplementedwith10%FBS(LifeTechnologies#26140-079),1%Penicillin/Streptomycin(Hyclone#SV30010.01)
CompleteGrowthMedium:ThawMedium2(BPSCat.#60184),1mg/mlofGeneticin(LifeTechnologies#11811031),and200μg/mlofHygromycinB(Hyclone#SV30070.01)Cellsshouldbegrownat37°Cwith5%CO2usingcompletegrowthmedium(ThawMedium2,Geneticin,andHygromycinB).
ApplicationReferences:
1. AstudyofPD-L1expressioninKRASmutantnon-smallcelllungcancercelllinesexposedtorelevanttargetedtreatments.(2017)
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