Pierce cypridina firefly luciferase dual assay kit

Manufactured by Thermo Fisher Scientific

The Pierce™ Cypridina-Firefly Luciferase Dual Assay Kit is a bioluminescent assay that measures the activity of two luciferase reporter enzymes, Cypridina and firefly luciferase, simultaneously in a single sample.

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Dual luciferase kit Firefly luciferase Luciferase assay

2 protocols using pierce cypridina firefly luciferase dual assay kit

Dual Luciferase Assay for NF-κB and IFNL Regulation

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Approximately 10000 HEK 293T cells were seeded in a 96-well plate and incubated for 16 to 18 h prior to plasmid transfection. Briefly, pMCS-Cypridina Luc Vector inserted with or without NF-κB and NS1 enriched locus, IFNL1, IFNL2, or IFNL3 with red firefly luc control plasmid were co-transfected into HEK 293T cells for 48 h. The primers for subcloning are listed in Supplementary Table S2. The p65 (OriGene, RG220780), p50 (OriGene, SC127485), and NS1 (OriGene, VC102470) plasmids were co-transfected with each pMCS-Cypridina Luc Vector with or without the above-enriched locus and red firefly luc control plasmid into HEK293 cells as described above. Luciferase activity was measured 24 h post-transfection using the TECAN Infinite 200 reader and Pierce™ Cypridina-Firefly Luciferase Dual Assay Kit (Thermo Scientific) according to the manufacturer’s protocol.

Lee M.C., Yu C.P., Chen X.H., Liu M.T., Yang J.R., Chen A.Y, & Huang C.H. (2022). Influenza A virus NS1 protein represses antiviral immune response by hijacking NF-κB to mediate transcription of type III IFN. Frontiers in Cellular and Infection Microbiology, 12, 998584.

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Dual Luciferase Assay for miR-103 and PTEN Interaction

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The luciferase reporter assay was performed according to the manufacturer’s instructions of Pierce™ Cypridina-Firefly Luciferase Dual Assay Kit (16184, Thermo). Briefly, AML-12 cells were co-transfected with a 10 nM miR-103 or NC control, a 2 ng pRL-CMV, and a 20 ng firefly luciferase reporter plasmid containing PTEN of the wild-type or mutant 30-untranslated region. Then, 48 h after transfection, the cell lysates were determined by luciferase to observe the interaction between miR-103 and PTEN. The liver tissues with the size of 1 mm × 1 mm × 1 mm were fixed, dehydrated, impregnated, and embedded to make ultrathin sections (50-70 nm) and then stained with uranium acetate and lead citrate successively and dried for observation under TEM.

Lu M.M., Ren Y., Zhou Y.W., Xu L.L., Zhang M.M., Ding L.P., Cheng W.X, & Jin X. (2023). Antagonizing adipose tissue-derived exosome miR-103-hepatocyte phosphatase and tensin homolog pathway alleviates autophagy in non-alcoholic steatohepatitis: A trans-cellular crosstalk. World Journal of Gastroenterology, 29(29), 4528-4541.

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