Luciferase Assay for ROB and DOC Treatment

PC-3/N cells³⁵ (link) were treated with ROB and/or DOC for 24 h, and then the cells were harvested in 1× reporter lysis buffer (Promega, Madison, WI, USA). After centrifugation, 10 µl aliquots of the supernatants were mixed with 10 µl of luciferase substrate (Promega) and measured for the luciferase activity by using a Luminometer LuMate™ (Awareness Technology, Palm City, FL, USA). The luciferase activity was normalised against known protein concentrations and expressed as the percentage of luciferase activity in the control cells. The protein level was determined by Bio-Rad protein assay kits (Bio-Rad, Hercules, CA, USA) according to the manufacturer's instructions.

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Wang X., Xuetao X., Wu M., Wu P., Sheng Z., Liu W., Ma Y.Y., Zhao D.G., Zhang K., Li D., Zheng X, & Goodin S. (2022). Inhibitory effect of roburic acid in combination with docetaxel on human prostate cancer cells. Journal of Enzyme Inhibition and Medicinal Chemistry, 37(1), 542-553.

Publication 2022

1× reporter lysis buffer luciferase substrate Bio-Rad protein assay kits

Assay Awareness Buffer Cells Centrifugation Luciferase Palm Promega Protein

Corresponding Organization : Wuyi University

Other organizations : Rutgers, The State University of New Jersey

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Variable analysis

independent variables

ROB (treatment)
DOC (treatment)

dependent variables

Luciferase activity (normalized against protein concentration, expressed as percentage of control)

control variables

Protein concentration (determined by Bio-Rad protein assay)

controls

Control cells (untreated)

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