AR negative cell lines (M12, PC3 and LNCaPAPIPC) (see the supplemental data for details) were co-transfected with the p3XFLAG-CMV10-based expression vector and the reporter plasmid. Specifically, the reporter assay was done with either the luciferase reporter driven by canonical androgen responsive regions (pARR2PB-luc)25 (link), or a luciferase construct under the control of AR-V-specific binding sites derived from the promoter element of the ubiquitin-conjugating enzyme E2C (UBE2C) gene (UBE2C–luc)26 (link), 27 (link), 59. Renilla luciferase expression plasmid was used as an internal standard in conjunction with pARR2PB-luc. At 16 h after transfection, cells were treated with 1 nM 5α-dihydrotestosterone (DHT) or vehicle control. As necessary, 10 µM enzalutamide was added 2 h prior to DHT treatment. Luciferase assay was done at 24 h after DHT treatment using the Dual-Glo luciferase assay system (Promega, Madison, WI, USA) with GloMax Multi Detection System for detection (Promega). Firefly luciferase luminescence was normalized to that of the co-expressed Renilla luciferase or protein concentration per each sample.
Glomax multi detection system for detection
Manufactured by Promega
Sourced in United States
The GloMax Multi Detection System is a versatile laboratory instrument designed for the detection and quantification of various luminescent and fluorescent signals. It offers a range of detection capabilities, including luminescence, fluorescence, and absorbance measurements, making it suitable for a wide array of applications in life science research.
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Lab products found in correlation
2 protocols using glomax multi detection system for detection
1
AR Transcriptional Regulation Assay
2
AR Signaling Pathway Regulation
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AR negative cell lines (M12, PC3 and LNCaPAPIPC) (see the Supplementary Data for details) were co-transfected with the p3XFLAG-CMV10-based expression vector and the reporter plasmid. Specifically, the reporter assay was done with either the luciferase reporter driven by canonical androgen responsive regions (pARR2PB-luc),23 or a luciferase construct under the control of AR-V-specific binding sites derived from the promoter element of the ubiquitin-conjugating enzyme E2C (UBE2C) gene (UBE2C-luc).24 (link), 25 (link), 58 (link)Renilla luciferase expression plasmid was used as an internal standard in conjunction with pARR2PB-luc. At 16 h after transfection, cells were treated with 1 nm 5α-dihydrotestosterone (DHT) or vehicle control. As necessary, 10 μm enzalutamide was added 2 h prior to DHT treatment. Luciferase assay was done at 24 h after DHT treatment using the Dual-Glo luciferase assay system (Promega, Madison, WI, USA) with GloMax Multi Detection System for detection (Promega). Firefly luciferase luminescence was normalized to that of the co-expressed Renilla luciferase or protein concentration per each sample.
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