Luc-Pair™ miR Luciferase Assay

TheLuc-Pair™ miRLuciferaseAssayKitprovides an efficientsystem in a convenient 96-wellplateformat formeasuringfireflyand Renilla luciferases sequentially.OptimizedforusewithGeneCopoeia miRNA3’ UTRTargetSequenceExpressionClones, Luc-PairmiRassays,enableseasy,costeffective validation of miRNAactivity.Luc-PairmiRLuciferase Assaysprovide the followingadvantages:
  • High sensitivity in a large dynamic range
  • Dual reporter system
  • Reproducible
  • Cost effectiveness
  • Simple assay format
  • Suitable for HTS
Figure1.The inhibitory effect of miRNA on a target sequence(3’ UTR) expression clonecan be measured with GeneCopoeia Luc-PairmiR Luciferase Assay kit. HEK 293 cellswereplated on a 6-wellplate. On the secondday,thecellsweretransfectedwith1.0 mg of targetsequence(3’ UTR)expressionclone(pEZX-MT01-Lin28 UTR-fLuc) and 1.4 mg of miRNA expressionvector(ormiRNAcontrolvector) as indicated in thefigure. Thecellsweretransferred to a 96-wellplate 18 hoursaftertransfection and culturedforanother 24 hours.Bothfireflyluciferase and Renilla luciferaseactivitiesweremeasured as describedanddatawasrecorded on Victor II machine.Fireflyluciferaseactivitywasthennormalizedwith Renilla luciferaseactivities in thesamewell.
   
Figure2. Linear relationship between emitted light and amount of luciferase expression vector transfected in HEK293. HEK293cellswere plated on a 6-wellplate. On thesecondday,thecellsweretransfected withdifferentamounts of GeneCopoeiapMRO or pEZX-MT01 miRNA3” UTRTargetSequenceClone.Thecellsweretransferred to a 96-wellplate 18 hoursaftertransfection and culturedforanother 24 hours.Firefly luciferaseassaywasthenperformed on pMROvector(Fig.2a) and Renilla luciferaseassaywasperformed on pEZX-MT01 vector(Fig.2b) as described.
 
Catalog No. Name Size
LPFR-M010 Luc-Pair™ miRLuciferase Assay Kit (1) 100 reactions
LPFR-M100 Luc-Pair™ miRLuciferase Assay Kit (10) 1000 reactions
 

Advantages

  • Ideal for GeneCopoeia miRNA 3’ UTR Target Sequence Expression Clones
  • Easy to use — less hands-on time and quick results
  • Suitable for micro-plate format — save time and money