Cells were transfected with HOP-Flash (Addgene#83467) luciferase reporter construct along with pRL-CMV-Renilla luciferase control reporter vector by Fugene6 (Roche). Luciferase activity was measured with Dual Luciferase Reporter Assay System (Promega; #E1910) according to manufacturer’s instructions. The reporter’s firefly luciferase activity was normalized to the levels of Renilla luciferase used as an internal control reporter. The relative luciferase activity displayed on the Y-axis indicates the ratio between Firefly/Renilla luciferase activities.
Hop flash
Manufactured by Addgene
The HOP-Flash is a high-performance liquid chromatography (HPLC) system designed for rapid and efficient purification of biomolecules. It features a compact and user-friendly design with intuitive software control. The HOP-Flash system is capable of performing analytical and preparative separations with high resolution and recovery.
Automatically generated - may contain errors
Lab products found in correlation
Prl tk renilla plasmid, by Promega (5 mentions)
Dual luciferase reporter assay kit, by Promega (5 mentions)
Hip flash luciferase reporter plasmid, by Addgene (4 mentions)
Lipofectamine 3000, by Thermo Fisher Scientific (4 mentions)
Dual luciferase reporter assay system, by Promega (1 mentions)
Neon transfection system, by Thermo Fisher Scientific (1 mentions)
Hip flash, by Addgene (1 mentions)
6 protocols using hop flash
1
Quantifying Transcriptional Regulation via Luciferase Assay
2
Luciferase Reporter Assay for Transcription Factor Activity
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Cells (4 × 104) were seeded in triplicate in 24-well plates and cultured for 24 h, and the luciferase reporter assay was performed as previously described.55 (link) Cells were transfected with 100 ng HOP-Flash (Cat. #83467; Addgene) or HIP-Flash luciferase reporter plasmid (Cat. #83466; Addgene), plus 5 ng pRL-TK Renilla plasmid (Promega) using Lipofectamine 3000 (Invitrogen) according to the manufacturer’s recommendation. Luciferase and Renilla signals were measured 36 h after transfection using a Dual Luciferase Reporter Assay Kit (Promega) according to the manufacturer’s protocol.
3
Luciferase Assay for Wnt Signaling
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Cells (4 × 104) were seeded in triplicate in 24-well plates and cultured for 24 h, and the luciferase reporter assay was performed as previously described (32 (link)). Cells were transfected with 100 ng HOP-Flash (Catalog # 83467, Addgene) or HIP-Flash luciferase reporter plasmid (Catalog # 83466, Addgene), plus 5 ng pRL-TK Renilla plasmid (Promega) using Lipofectamine 3000 (Invitrogen) according to the manufacturer’s recommendation. Luciferase and Renilla signals were measured 36 h after transfection using a Dual Luciferase Reporter Assay Kit (Promega) according to the manufacturer’s protocol.
4
Luciferase Assay for Transcriptional Activity
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Cells (4 × 104) were seeded in triplicate in 24-well plates and cultured for 24 h, and the luciferase reporter assay was performed as previously described (47 (link)). Cells were transfected with 100 ng HOP-Flash (Catalog # 83467, Addgene) or HIP-Flash luciferase reporter plasmid (Catalog # 83466, Addgene), plus 5 ng pRL-TK Renilla plasmid (Promega) using Lipofectamine 3000 (Invitrogen) according to the manufacturer’s recommendation. Luciferase and Renilla signals were measured 36 h after transfection using a Dual Luciferase Reporter Assay Kit (Promega) according to the manufacturer’s protocol.
5
Dual Luciferase Reporter Assay for Wnt Signaling
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Cells (4 × 104) were seeded in triplicate in 24-well plates and cultured for 24 h, and the luciferase reporter assay was performed as previously described.35 Cells were transfected with 100 ng HOP-Flash (83467, Addgene) or HIP-Flash luciferase reporter plasmid (83466, Addgene), plus 5 ng pRL-TK Renilla plasmid (Promega) using Lipofectamine 3000 (Invitrogen), according to the manufacturer’s recommendation. Luciferase and Renilla signals were measured 36 h after transfection using a Dual Luciferase Reporter Assay Kit (Promega), according to the manufacturer’s protocol.
6
Polaprezinc and ZnSO4 Modulation of Luciferase Activity
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Cells were seeded in triplicate in six-well culture plates at 1 × 105 cells and cultured for 24 h, and the luciferase reporter assay was performed as previously described53 . The cells were transfected with 500 ng of HOP-Flash (83467, Addgene) or HIP-Flash luciferase reporter plasmid (83466, Addgene) along with 10 ng of pRL-TK Renilla plasmid (Promega) using the Neon Transfection System (Invitrogen) according to the manufacturer’s instructions. The day after transfection, cells were treated with 50 μM polaprezinc or ZnSO4, respectively. Luciferase and Renilla signals were measured 48 h after transfection using a Dual Luciferase Reporter Assay Kit (Promega, Madison, WI, USA) according to the manufacturer’s instructions.
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