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Renilla-Firefly Luciferase Dual Assay Kit

40 Cited Publications
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Based on 40 publication(s) in Google Scholar

MCE Renilla-Firefly Luciferase Dual Assay Kit is designed to be used for high-throughput, rapid quantitation of both Firefly and Renilla luciferases from a single sample in mammalian cell culture.

  • Storage :
    -20℃, 1 year. Protecting from light.
  • Experimental Validation Results for Renilla-Firefly Luciferase Dual Assay Kit
  • Experimental Validation Results for Renilla-Firefly Luciferase Dual Assay Kit
  • Experimental Validation Results for Renilla-Firefly Luciferase Dual Assay Kit
  • Experimental Validation Results for Renilla-Firefly Luciferase Dual Assay Kit
  • Experimental Validation Results for Renilla-Firefly Luciferase Dual Assay Kit
  • Renilla-Firefly Luciferase Dual Assay Kit Product Image
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To place orders, for customer services and technical support, please contact: MedChemExpress USA

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Description & Advantages

MCE Renilla-Firefly Luciferase Dual Assay Kit contains high-purity D-Luciferin, coelenterazine and proprietary reaction buffer. The Firefly Luciferase Working Solution is first added to the sample. This reagent contains D-Luciferin which can produce Firefly luciferase bioluminescence. Next, the Renilla Luciferase Working Solution is added to the same sample. It quenches the Firefly luciferase bioluminescence and produce Renilla luciferase bioluminescence. The light production of both reactions can be conveniently measured on a luminometer.

Protocol

1. Material Preparation

1.1 Preparation of Lysis Buffer (1×)

Dilute Lysis Buffer (5×) 1:5 with dH2O (e.g. 1 mL Lysis Buffer + 4 mL dH2O)

1.2 Preparation of Firefly Luciferase Working Solution

Briefly centrifuge the Firefly Luciferase Substrate before opening the tube. Add 1 mL of Firefly Luciferase Buffer and mix thoroughly. Transfer the entire solution to the remaining 9 mL of Firefly Luciferase Buffer and mix well to obtain the working solution.

1.3 Preparation of Renilla Luciferase Working Solution

Briefly centrifuge the Renilla Luciferase Substrate before opening the tube. Add 1 mL of Renilla Luciferase Buffer and mix thoroughly. Transfer the entire solution to the remaining 9 mL of Renilla Luciferase Buffer and mix well to obtain the working solution.

Note: Firefly Luciferase Working Solution and Renilla Luciferase Working Solution can be dispensed into aliquots. When protecting from light and avoiding repetitive freeze-thaw cycles, the reagent is stable for one month at -20°C or for one year when stored at -80°C.

2. Cell Lysis

2.1 Determine transfection parameters (plated cell density and subsequent incubation time) to ensure that cells are no more than 95% confluent at the desired time of lysate preparation.

2.2 Carefully remove culture medium from cells. Wash cells with wash solution, such as PBS, normal saline orserum-free culture medium, to remove residual medium.

2.3 Add proper volume of Lysis Buffer (1×). The recommended volumes of Lysis Buffer (1×) to add per well are as follows:

Multi-well Plate Lysis Buffer/Well
6-well 500 μL
12-well 200 μL
24-well 100 μL
48-well 50 μL
96-well 30 μL

Note: The volume of added Lysis Buffer (1×) can be adjusted proportionally.

2.4 Add proper volume of Lysis Buffer (1×). The recommended volumes of Lysis Buffer (1×) to add per well are as follows:

2.5 Place the culture plates on a shaker with gentle shaking at room temperature for 15 minutes.

2.6 After lysis, centrifuge at 12,000 rpm for 10 minutes at 4 °C. Transfer the supernatants to a new tube for further analysis.

3. Luciferase Dual Assay

3.1 Add 20μL of cell lysate to a black flat-bottomed 96-well plate.

3.2 Add 100 μL of Firefly Luciferase Working Solution into each well containing cell lysate.

3.3 Immediately after adding the reagent, read the sample to capture the Firefly luciferase signals.

3.4 Add 100 μL of Renilla Luciferase Working Solution into each well from step 3.3, then immediately read the sample again to capture the Renilla luciferase signals.

3.5 Calculate the ratio of luminescence from the Firefly luciferase to the Renilla luciferase.

Note: 1) Assure Firefly Luciferase Working Solution and Renilla Luciferase Working Solution are equilibrated to room temperature before use.

2) The luminescence values of Firefly and Renilla Luciferase should be subtracted from the corresponding background values.

Storage

-20℃, 1 year.

Protecting from light.

Components

Components HY-K1013-100T
Firefly Luciferase Buffer 10 mL
Firefly Luciferase Substrate 1 vial
Renilla Luciferase Buffer 10 mL
Renilla Luciferase Substrate 1 vial
Lysis Buffer (5×) 10 mL

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  • Experimental Validation Results for Renilla-Firefly Luciferase Dual Assay Kit

    Renilla-Firefly Luciferase Dual Assay Kit purchased from MedChemExpress. Usage Cited in: Cell Discov. 2026 Jan 20;12(1):5. [Abstract]

    Luciferase activities of WT or Mut BCAT1 and GLS reporter vectors were quantified in I-hASCs under defined genetic perturbations: control, IGF2BP3 overexpression (IGF2BP3), or IGF2BP3 overexpression with METTL3 knockdown (IGF2BP3+shMETTL3). Luciferase activity was quantified using the Renilla-Firefly Luciferase Dual Assay Kit.

  • Experimental Validation Results for Renilla-Firefly Luciferase Dual Assay Kit

    Renilla-Firefly Luciferase Dual Assay Kit purchased from MedChemExpress. Usage Cited in: Nat Commun. 2025 Apr 28;16(1):3975. [Abstract]

    The relative activity of the Anxa1 promoter was detected via luciferase assay in three groups. All experiments were independently repeated in triplicate. Luciferase activity results showed that Sox9 overexpression significantly activated Anxa1 gene transcription in Moc1 cells transfected with the reporter construct. Motif deletion abolished this induction of the Anxa1 promoter. The Renilla-Firefly Luciferase Dual Assay Kit was used for the detection and analysis of luciferase activity.

  • Experimental Validation Results for Renilla-Firefly Luciferase Dual Assay Kit

    Renilla-Firefly Luciferase Dual Assay Kit purchased from MedChemExpress. Usage Cited in: Cell Rep Med. 2025 Jun 17;6(6):102146. [Abstract]

    Luciferase reporter assay to confirm the transcriptional activation of ADAM10 by DDX5 and E2F1 (3 independent experiments). Luciferase activity was quantified using the Renilla-Firefly Luciferase Dual Assay Kit.

  • Experimental Validation Results for Renilla-Firefly Luciferase Dual Assay Kit

    Renilla-Firefly Luciferase Dual Assay Kit purchased from MedChemExpress. Usage Cited in: J Transl Med. 2025 Apr 2;23(1):387. [Abstract]

    Luciferase activity was quantified using the Renilla-Firefly Luciferase Dual Assay Kit.
    Relative luciferase activity. **P < 0.05 vs. Group (TGF-β1+, miR-223-3p-, CHUK plasmid-). **** P < 0.01 vs. Group (TGF-β1+, miR-223-3p-, CHUK plasmid-). The results of double luciferase reporter gene experiment showed that the relative activity of luciferase decreased significantly (P<0.01) after co-transfection of miR-223-3p mimics and wild-type CHUK vector, while co-transfected miR-223-3p mimics and mutant CHUK vectors showed no significant effect on the relative activity of luciferase (P>0.05), indicating that miR-223-3p could target IKKα mRNA to inhibit downstream NF-κB signaling pathway and suppress the EMT of HK2 cells.

  • Experimental Validation Results for Renilla-Firefly Luciferase Dual Assay Kit

    Renilla-Firefly Luciferase Dual Assay Kit purchased from MedChemExpress. Usage Cited in: J Clin Invest. 2023 Oct 16;133(20):e170173. [Abstract]

    Luciferase activity of TET2 5′-UTR WT or TET2 5′-UTR m6Am site mutation (5′-UTR MUT) in SCC1 control cells and cells transfected with WT (OE) or mutant PCIF1 plasmid (OEmut) (n = 3). Luciferase activity was quantified using the Renilla-Firefly Luciferase Dual Assay Kit.

Assay Types
  • Experimental Validation Results for Renilla-Firefly Luciferase Dual Assay Kit
    1/16

    Renilla-Firefly Luciferase Dual Assay Kit purchased from MedChemExpress. Usage Cited in: Cell Discov. 2026 Jan 20;12(1):5. [Abstract]

    Luciferase activities of WT or Mut BCAT1 and GLS reporter vectors were quantified in I-hASCs under defined genetic perturbations: control, IGF2BP3 overexpression (IGF2BP3), or IGF2BP3 overexpression with METTL3 knockdown (IGF2BP3+shMETTL3). Luciferase activity was quantified using the Renilla-Firefly Luciferase Dual Assay Kit.

  • Experimental Validation Results for Renilla-Firefly Luciferase Dual Assay Kit
    1/16

    Renilla-Firefly Luciferase Dual Assay Kit purchased from MedChemExpress. Usage Cited in: Nat Commun. 2025 Apr 28;16(1):3975. [Abstract]

    The relative activity of the Anxa1 promoter was detected via luciferase assay in three groups. All experiments were independently repeated in triplicate. Luciferase activity results showed that Sox9 overexpression significantly activated Anxa1 gene transcription in Moc1 cells transfected with the reporter construct. Motif deletion abolished this induction of the Anxa1 promoter. The Renilla-Firefly Luciferase Dual Assay Kit was used for the detection and analysis of luciferase activity.

  • Experimental Validation Results for Renilla-Firefly Luciferase Dual Assay Kit
    1/16

    Renilla-Firefly Luciferase Dual Assay Kit purchased from MedChemExpress. Usage Cited in: Cell Rep Med. 2025 Jun 17;6(6):102146. [Abstract]

    Luciferase reporter assay to confirm the transcriptional activation of ADAM10 by DDX5 and E2F1 (3 independent experiments). Luciferase activity was quantified using the Renilla-Firefly Luciferase Dual Assay Kit.

  • Experimental Validation Results for Renilla-Firefly Luciferase Dual Assay Kit
    1/16

    Renilla-Firefly Luciferase Dual Assay Kit purchased from MedChemExpress. Usage Cited in: J Transl Med. 2025 Apr 2;23(1):387. [Abstract]

    Luciferase activity was quantified using the Renilla-Firefly Luciferase Dual Assay Kit.
    Relative luciferase activity. **P < 0.05 vs. Group (TGF-β1+, miR-223-3p-, CHUK plasmid-). **** P < 0.01 vs. Group (TGF-β1+, miR-223-3p-, CHUK plasmid-). The results of double luciferase reporter gene experiment showed that the relative activity of luciferase decreased significantly (P<0.01) after co-transfection of miR-223-3p mimics and wild-type CHUK vector, while co-transfected miR-223-3p mimics and mutant CHUK vectors showed no significant effect on the relative activity of luciferase (P>0.05), indicating that miR-223-3p could target IKKα mRNA to inhibit downstream NF-κB signaling pathway and suppress the EMT of HK2 cells.

  • Experimental Validation Results for Renilla-Firefly Luciferase Dual Assay Kit
    1/16

    Renilla-Firefly Luciferase Dual Assay Kit purchased from MedChemExpress. Usage Cited in: J Clin Invest. 2023 Oct 16;133(20):e170173. [Abstract]

    Luciferase activity of TET2 5′-UTR WT or TET2 5′-UTR m6Am site mutation (5′-UTR MUT) in SCC1 control cells and cells transfected with WT (OE) or mutant PCIF1 plasmid (OEmut) (n = 3). Luciferase activity was quantified using the Renilla-Firefly Luciferase Dual Assay Kit.

  • Renilla-Firefly Luciferase Dual Assay Kit Product Image
    1/16
  • Experimental Validation Results for Renilla-Firefly Luciferase Dual Assay Kit
  • Experimental Validation Results for Renilla-Firefly Luciferase Dual Assay Kit
  • Experimental Validation Results for Renilla-Firefly Luciferase Dual Assay Kit
  • Experimental Validation Results for Renilla-Firefly Luciferase Dual Assay Kit
  • Experimental Validation Results for Renilla-Firefly Luciferase Dual Assay Kit
  • Renilla-Firefly Luciferase Dual Assay Kit Product Image