To confirm miR-129-5p binding to SOX4 3′UTR we used luciferase reporter clone for human SOX4 3′UTR (HmiT017630-MT06, Genecopoeia) and luciferase control reporter construct (CmiT000001-MT06, Genecopoeia). Constructs were transfected in A375 (ctrl) and A375 miR129-5p (overexpression) cells 24 h after seeding. The relative Luciferase activity was measured 48 h after transfection using Luc-Pair Duo-Luciferase Assay Kits 2.0 (Genecopoeia) following the manufacturer’s instructions.
Luc pair duo luciferase assay kits 2
Manufactured by GeneCopoeia
Sourced in United States
The Luc-Pair Duo-Luciferase Assay Kits 2.0 are a set of reagents designed to measure the activity of two different luciferase reporter genes simultaneously. The kit includes a Firefly Luciferase Assay Reagent and a Renilla Luciferase Assay Reagent, which can be used to quantify the activity of Firefly and Renilla luciferase reporter enzymes, respectively.
Automatically generated - may contain errors
Lab products found in correlation
Cmit000001 mt06, by GeneCopoeia (1 mentions)
Lipofectamine ltx with plus reagent, by Thermo Fisher Scientific (1 mentions)
Lipofectamine rnaimax, by Thermo Fisher Scientific (1 mentions)
Pezx fr02 vector, by GeneCopoeia (1 mentions)
Microplate reader, by Molecular Devices (1 mentions)
Lipofectamine 3000, by Thermo Fisher Scientific (1 mentions)
5 protocols using luc pair duo luciferase assay kits 2
1
Validating miR-129-5p Regulation of SOX4 via Luciferase Assay
2
Characterization of Maf and Il23r 3' UTR Interaction with miR-221/222
3
Dual-Luciferase Assay of USP37 3'-UTR
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WT or mutant fragments of 3′-untranslated region (3′-UTR) of USP37 were amplified and inserted downstream of the firefly luciferase reporter gene in the pEZX-FR02 vector (Genecopoeia, Rockville, MD). Next, 100 nM mimic of miR-4487 and 250 ng pEZX-EF02 vectors containing WT and mutated USP37 were co-transfected into HEK293 cells using lipofectamine RNAiMAX and 3000 reagents, respectively (Life Technologies Inc.). After an additional cell incubation of 48 hours, the luciferase activity (Fluc/Rluc) was assayed using the Luc-Pair Duo-Luciferase Assay Kits 2.0 (Genecopoeia).
4
BRCA1 Promoter Luciferase Assay
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The construction of Sei-1 vectors (EX-A3731-Lv103) and pGL3-BRCA1-promoter luciferase reporter vectors were completed by GeneCopoeia company (Guangzhou, China) and Aizhe Biological Technology Co., Ltd (Guangzhou, China), respectively. The 293T cells were plated in 24-well plates at 50,000/well. The co-transfection of Sei-1 and luciferase reporter vectors were accomplished according to the instruction of HilyMax reagents (Dojindo, Shanghai, China). The detection of luciferase activity was based on the instruction of Luc-Pair Duo-Luciferase Assay Kits 2.0 (GeneCopoeia, Rockville, MD, USA) and completed by microplate reader (Molecular Devices, Shanghai, China).
5
Dual-Luciferase Assay for miRNA Targeting
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The dual-luciferase constructs were purchased from Genecopoeia: miR-200b 3’UTR #MmiT073700-MT06 optimized for miRNA targeting at Genbank: NM_001159517.1 3’UTR and control scramble 3’UTR # CmiT000001-MT06. Briefly, the plasmids were transfected using Lipofectamine 3000 (Invitrogen, L3000015) and incubated for 48 h at 37°C with 5% CO2. Cells were lysed and luciferase activity was assessed by Luc-Pair Duo-Luciferase Assay Kits 2.0 (Genecopoeia, LF001). Renilla luciferase activity was normalized to the corresponding firefly luciferase activity and plotted as a percentage of the control.
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