A dual-luciferase reporter gene assay (Promega, Madison, WI) was used to determine promoter activity. Briefly, cells were plated in 12-well plates and transiently transfected with 1 μg/ml reporter plasmids and phRL-SV plasmid (hRenilla luciferase expression for normalization) using Hillymax® reagent (Dojindo Molecular Technologies, Gaithersburg, MD). The cells were then incubated in culture medium without serum. Firefly and hRenilla luciferase activities in the cell lysates were measured using a luminometer (LB941, Berthold Technologies, Bad Wild, Germany). Relative luciferase activities were calculated by normalizing the promoter-driven firefly luciferase activity to the hRenilla luciferase.
Hillymax reagent
Manufactured by Dojindo Laboratories
Hillymax® reagent is a laboratory product manufactured by Dojindo Laboratories. It is a chemical reagent designed for use in various scientific applications. The core function of the Hillymax® reagent is to facilitate specific chemical reactions or analyses, though the precise intended use may vary depending on the application.
Automatically generated - may contain errors
3 protocols using hillymax reagent
1
Dual-Luciferase Reporter Assay for Promoter Activity
2
Investigating KIF2A Silencing in AML Cells
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The transfection of 50 nM negative control (NC) or KIF2A siRNA (both Shanghai GenePharma Co., Ltd.) into AML cells (KG-1 and Kasumi-1) was performed using HillyMax reagent (Dojindo Molecular Technologies, Inc.) at 37°C for 6 h. Untreated cells were labeled as Untreated cells. At 48 h after transfection, RT-qPCR, western blotting and Annexin V/propidium iodide (AV/PI) and chemosensitivity assay were performed. At 0, 24, 48 and 72 h after transfection, Cell Counting Kit-8 (CCK-8) assay was performed. The target sequence of KIF2A siRNA was 5′-GGCAAAGAGAUUGACCUGG-3′ and the scrambled sequence used for NC siRNA was 5′-AAGAACAACACAAAAGAACAG-3′.
3
Dual-Luciferase Reporter Assay for Promoter Activity
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
A dual-luciferase reporter gene assay system (Promega, Madison, WI) was used to determine promoter activity. Briefly, cells were plated in 12-well plates and transiently transfected with 1 μg/ml reporter plasmids and phRL-SV plasmid (hRenilla luciferase expression for normalization) using Hillymax® reagent (Dojindo Molecular Technologies, Gaithersburg, MD). The cells were then incubated in culture medium without serum for 18 h. Firefly and hRenilla luciferase activities in the cell lysates were measured using a luminometer (LB941, Berthold Technologies, Bad Wild, Germany). Relative luciferase activities were calculated by normalizing the promoter-driven firefly luciferase activity to the hRenilla luciferase.
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