PolarScreen AR Competitor and PolarScreen GR Competitor assays (Green kit; Invitrogen) were used to measure the binding affinities of the compounds for the AR and GR, according to manufacturer instructions, respectively. The preservatives were tested at concentrations from down to (in dilutions steps of 1:10) in both assays, with down to (in dilutions steps of 1:10) dihydrotestosterone as control ligand for AR and down to (in dilutions steps of 1:10) dexamethasone as control ligand for GR. The fluorescence polarization was recorded using a microplate reader (Synergy 4 Hybrid Multi-Mode; BioTek).
Synergy 4 hybrid multi mode
Manufactured by Agilent Technologies
Sourced in Canada
The Synergy 4 Hybrid Multi-Mode is a multi-mode microplate reader that can perform absorbance, fluorescence, and luminescence measurements. It is designed to provide flexible and efficient performance for various applications in life science research and drug discovery.
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Lab products found in correlation
3 protocols using synergy 4 hybrid multi mode
1
Competitive Binding Assays for AR and GR
2
Luciferase Inhibition Assay for T3 Response
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For the luciferase inhibition assay, GH3.TRE-Luc cells in were seeded in 96-well plates and incubated for 24 h with T3, followed by cell lysis using Luciferase Cell Culture Lysis Reagent (Promega). Serial dilutions for each treatment were prepared in the cell lysate and incubated at room temperature for 30 min. Then firefly luciferase reagent ONE-Glo (Promega) was added, and luciferase fluorescence was recorded (2-s medium shaking step followed by luminescence end point measurement; no light source or emission filters) using a microplate reader (Synergy 4 Hybrid Multi-Mode; BioTek).
3
MTT Assay for Cell Viability
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Cell viability was determined using 3-(4, 5-dimethylthiazal-2-yl)-2, 5-diphenyl-tetrazolium bromide (MTT) assay as previously described [22] . Briefly, SRA01/04 cells were plated onto 96-well plates at a density of 1 × 10 4 cells/well in 0.1 ml of DMEM medium. After the transfection of siRNA-H19, miR-675 inhibitor, miR-675 mimic, or their negative controls for 48 h, these cells were incubated with MTT at a final concentration of 0.5 mg/ml for 4 h at 37°C. The formazan crystals were dissolved by adding 100 mM DMSO after medium removal. Absorbance was measured using a microplate reader (Synergy 4 Hybrid Multi-Mode; BioTek Instruments, Canada).
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