The promoter regions of TRPC1 were cloned into a PGL3 luciferase reporter vector (Addgene, Cambridge, Mass, USA). The vector was transfected into a 24-well plate. After 48 h, the cells were lysed in 100 μL of lysis buffer and then assayed for promoter activity using a dual-luciferase reporter assay system (Promega, Madison, WI, USA). The Renilla luciferase signal was normalized to the firefly luciferase signal. The enzymatic activity of luciferase was measured using a PerkinElmer 2030 Multilabel Reader (PerkinElmer, Waltham, MA, USA).
Pgl3 luciferase reporter vector
Manufactured by Addgene
Sourced in United States
The PGL3 luciferase reporter vector is a plasmid designed for the expression and measurement of luciferase activity in mammalian cells. The vector contains the firefly luciferase gene under the control of a promoter, allowing for the quantification of gene expression levels.
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Lab products found in correlation
2 protocols using pgl3 luciferase reporter vector
1
Quantifying Promoter Activity of TRPC1 Gene
2
Assaying PDLIM5 Promoter Activity
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The PDLIM5 promoter region was cloned into the PGL3 luciferase reporter vector (Addgene, Cambridge, MA, USA). Then, the PDLIM5 promoter luciferase construct was transfected into cells in a 24-well plate for 48 h. Subsequently, the cells were lysed using 100 μL lysis buffer, and promoter activity was assayed using a dual luciferase reporter assay system (Promega, Madison, WI, USA). The luciferase activities were measured using a PerkinElmer 2030 Multilabel Reader (PerkinElmer, Waltham, MA, USA). The primers used are listed in Supplementary Table S3 .
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