Dual luciferase kit

Manufactured by Beyotime

Sourced in China

The Dual Luciferase Kit is a laboratory tool designed to measure the activity of two different luciferase reporter genes simultaneously. The kit provides reagents and protocols to quantify the activities of firefly and Renilla luciferase in a single sample, allowing for normalization and comparative analysis.

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Lab products found in correlation

Lipofectamine 2000, by Thermo Fisher Scientific (1 mentions) Gw9662, by Cayman Chemical (1 mentions) Rosiglitazone, by Cayman Chemical (1 mentions) Lipofectamine 3000 reagent, by Thermo Fisher Scientific (1 mentions) Dual luciferase reporter system, by Promega (1 mentions) Pmirglo vector, by Promega (1 mentions)

Spelling variants of this product

Dual Luciferase Reporter Gene Assay Kit (282 citations) Dual-luciferase reporter assay kit (115 citations) Dual-luciferase assay kit (51 citations) Dual-Lumi™ Luciferase Reporter Gene Assay Kit (26 citations) Dual luciferase reporter kit (22 citations) Dual-luciferase reporter gene detection kit (14 citations) Dual-Luciferase Detection kit (4 citations) Dual Luciferase Reporter Gene Assay Kit (RG027) (3 citations) Dual-specific luciferase assay kit (3 citations) Dual luciferase reporter gene test kit (2 citations)

7 protocols using dual luciferase kit

Gal 3'UTR miR-18a-5p Interaction Assay

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The predicted wild and mutated sequence of about 200 bp upstream and downstream of the interaction site with rno-miR-18a-5p (only mutation predicted interaction site) of the Gal 3′UTR was synthesized and was inserted into the luciferase reporter gene vector pmirGLO. HEK293T cells were transfected with the miR-18a-5p mimics or control along with the luciferase constructs using the transfection reagent Lipofectamine 2000 (Invitrogen). 48 hours after transfection, cells were lysed and chemiluminescence readings were taken with the Dual Luciferase Kit (Beyotime) and the instrument. Renilla luciferase was used as an internal reference, and the RLU value measured by firefly luciferase is divided by the RLU value measured by Renilla luciferase.

Xiao K., Song L., Hu Y., He W., Hou F., Yan P., Xu J., Wang K., Tao Y., Li D, & Xie L. (2021). Novel Role of miR-18a-5p and Galanin in Rat Lung Ischemia Reperfusion-Mediated Response. Oxidative Medicine and Cellular Longevity, 2021, 6621921.

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Binding efficiency of miRNA-mRNA pairs

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According to the miRwalk database, miR-675-5p binds to GSDME, and miR-1247-5p binds to HMGB1.To determine the binding efficiency between miR-675-5p and GSDME, miR-1247-5p and HMGB1, SH-SY5Y cells were co-transfected with miR-675-5p/miR-1247-5p mimics or mimics-NC (Sangon, China) with a luciferase construct containing GSDME/HMGB1 having wild-type binding site (Tsingke, China). Luciferase activities were analyzed using the Firefly Luciferase Reporter Gene Assay Cell Lysis Buffer (Beyotime, China; Cat No RG126S) and Dual Luciferase kit (Beyotime, China; Cat No RG088S) according to the manufacturer’s instructions.

Liang J., Wan Z., Qian C., Rasheed M., Cao C., Sun J., Wang X., Chen Z, & Deng Y. (2024). The pyroptosis mediated biomarker pattern: an emerging diagnostic approach for Parkinson’s disease. Cellular & Molecular Biology Letters, 29, 7.

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Dual-Luciferase Assay in A2780 and SKOV3

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A2780 and SKOV3 cells were transfected with the indicated plasmids (Supplemental methods). The firefly and renilla luciferase activities were measured using the Dual-Luciferase Kit (RG028, Beyotime, Shanghai, China) according to the manufacturer’s instructions.

Pan B., Wan T., Zhou Y., Huang S., Yuan L., Jiang Y., Zheng X., Liu P., Xiang H., Ju M., Luo R., Jia W., Lan C., Li J, & Zheng M. (2023). The MYBL2–CCL2 axis promotes tumor progression and resistance to anti-PD-1 therapy in ovarian cancer by inducing immunosuppressive macrophages. Cancer Cell International, 23, 248.

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Validation of miR-124-3p and STAT3 Interaction

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The predicted binding site fragments (miR-124-3p with STAT3) and mutation fragments were inserted into the dual-luciferase reporter vector as the reporter plasmids, which were labeled as WT and MUT, respectively. Luciferase reporter vectors were co-transfected with miR-124 mimic or mimic NC (Ribobio technology, Guangzhou, China). Next, 293T cells were seeded overnight, then co-treated with miR-124-3p reagents as well as the recombinant WT/MT-vector. Renilla fluorescence served as the internal reference. The cells were collected following the protocol of a dual-luciferase kit (Beyotime, Shanghai, China). The luciferase activity was detected using a PerkinElmer EnSpire Microplate reader (Promega Corporation, Madison, WI, USA).

Hong Y., Wang Y., Cui Y., Pan J., Mao S., Zhu Y., Wen T., Qi T., Wang A, & Luo Y. (2023). MicroRNA-124-3p Attenuated Retinal Neovascularization in Oxygen-Induced Retinopathy Mice by Inhibiting the Dysfunction of Retinal Neuroglial Cells through STAT3 Pathway. International Journal of Molecular Sciences, 24(14), 11767.

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Ppar-gamma Mediated Transcriptional Regulation

Cited 1 time

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Reporter constructs, pIDE and pIDE_ΔP, were generated by ligating varying lengths of IDE upstream sequence into the pGL₃-Basic luciferase expression vector (Hanheng Biotechnology Co., Shanghai, China) (Supplementary Figure 1). Lipofectamine 3000 Reagent (Invitrogen Life Technologies, California, USA) was used to transfect reporter constructs into HK-2 cells, as described in our previous report.^{[29 (link)]} At about 6 h post-transfection, the same amount of complete medium was added and then the cells were treated with 10 μM rosiglitazone, a selective PPARγ agonist (Cayman Chemical, Ann Arbor, Michigan), 5 μM GW9662, a PPARγ antagonist (Cayman Chemical, Ann Arbor, Michigan), or solvent for 42 h. Firefly and Renilla luciferase activities were measured using the Dual Luciferase Kit (Beyotime, Shanghai, China). The firefly luciferase values of each sample were normalized by Renilla luciferase activity and reported as relative light units.

Lawson A.J., Logan J.M., O'neill G.L., Desai M, & Stanley J. (1999). Large-Scale Survey of Campylobacter Species in Human Gastroenteritis by PCR and PCR–Enzyme-Linked Immunosorbent Assay. Journal of Clinical Microbiology, 37(12), 3860-3864.

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Dual Luciferase Assay for Analyzing Transcriptional Activity

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HEK 293T cells, 5 × 10⁴ well/500 μL, were seeded and cultured overnight in 24-well plate. Then the cells were transfected with either wild-type or mutant reporter gene plasmid using Lipo3000 reagent according to the manufacturer’s instructions. After 48 h, the cell lysates were collected and the activities of firefly and Renilla luciferases were measured by a Dual-Luciferase Reporter System (Promega). The dual luciferase kit (Beyotime, RG027) was used to measure luciferase activity. Relative luciferase activity was standardized by the Renilla luciferase. Each experiment was repeated three times.

Zhai Z., Ren Y., Shu C., Chen D., Liu X., Liang Y., Li A, & Zhou J. (2022). JAC1 targets YY1 mediated JWA/p38 MAPK signaling to inhibit proliferation and induce apoptosis in TNBC. Cell Death Discovery, 8, 169.

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GREM2 3'UTR Luciferase Assay

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Cells were transfected with pmir-GLO vector (Promega, Madison, WI, USA) including wild-type/mutant GREM2 3'UTR, miR-103a-3p mimics/NC-miRNA. They were transfected for 48 h and then detected via dual-luciferase kit (Beyotime Institute of Biotechnology, Shanghai, China).

Zhang Z, & Zhu X. (2022). MiR-103a-3p Contributes to the Progression of Colorectal Cancer by Regulating GREM2 Expression. Yonsei Medical Journal, 63(6), 520-529.

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