Luc-Pair™ miR Luciferase Assay Kit — optimized luciferase reporter assay for use with
GeneCopoeia miRNA Target Sequence 3' UTR Expression Clones
The Luc-Pair™ miR Luciferase Assay Kit provides an efficient system in a convenient 96-well plate format for measuring firefly and Renilla luciferases sequentially. Optimized for use with GeneCopoeia miRNA 3' UTR Target Sequence Expression Clones, Luc-Pair™ miR assays, enable easy, cost effective validation of miRNA activity.
Luc-Pair™ miR Luciferase Assays provide the following advantages:
- High sensitivity in a dynamic range
- Successive reporter system
- Consistent reproducibility
- Cost effectiveness
- Simple assay format
- Suitable for high-throughput screening
Measure inhibitory effect of a miRNA on a target sequence
Figure 1. The inhibitory effect of miRNA on a target sequence (3' UTR) expression clone can be measured with GeneCopoeia Luc-Pair™ miR Luciferase Assay kit. HEK 293 cells were plated on a 6-well plate. On the second day, the cells were transfected with 1.0 µg of Lin28 3' UTR miRNA target sequence expression clone in pEZX-MT01 vector with fLuc (Product ID: HmiT019205) and 1.4 µg of miR-125a precursor miRNA expression clone in pEZX-MR03 vector with eGFP (Product ID: HmiR0309) and miRNA control clone as indicated in the figure. The cells were transferred to a 96-well plate 18 hours after transfection and cultured for another 24 hours. Both firefly luciferase and Renilla luciferase activities were measured and data was recorded on Victor II machine. Firefly luciferase activity was then normalized with Renilla luciferase activities in the same well.
Linear range
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| Figure 2a. Firefly Luciferase Activity |
Figure 2b. Renilla Luciferase |
Figure 2. Linear relationship between emitted light and amount of luciferase expression vector transfected in HEK293. HEK 293 cells were plated on a 6-well plate. On the second day, the cells were transfected with different amounts of GeneCopoeia pMRO or pEZX-MT01 miRNA 3' UTR Target Sequence Clone as indicated in the figure. The cells were transferred to a 96-well plate 18 hours after transfection and cultured for another 24 hours. Firefly luciferase assay was then performed on pMRO vector (Fig. 2a) and Renilla luciferase assay was performed on pEZX-MT01 vector (Fig. 2b).
The advantages are clear
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