Peptide/SSO complexes were formed at MR5 using three different peptides: hPep1, hPep2, and hPep3. The complexes were then transferred to black 96-well plates containing SYBR™ Gold dye (Invitrogen™, Waltham, MA, USA) to measure the amount of accessible SSO in the complexes with SynergyMX fluorometer (BioTek, Winooski, VT, USA). For complex stability evaluation, competitive anion binder hexametaphosphate (HMP) was added at different concentrations from 0.04 to 40 µg/mL and the amount of released oligonucleotide was measured after the HMP addition. Measurements were done in duplicates and results show the mean of three independent experiments (mean ± SEM, n = 3).
Synergy mx fluorometer
Manufactured by Agilent Technologies
Sourced in United States, United Kingdom
The Synergy MX fluorometer is a high-performance microplate reader designed for a wide range of fluorescence-based assays. It features a unique optical system that enables sensitive detection and accurate quantification of fluorescent signals in multi-well plates.
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Lab products found in correlation
3 protocols using synergy mx fluorometer
1
Peptide-SSO Binding and Stability
2
Fluorometric Assay for DV2 RdRp Activity
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The experimental procedures were performed in 96-well, black-bottom microplate. The reaction mixture for measuring RdRp activity was prepared by mixing 3 pmol of RNA substrate, 3.5 pmol of DV2 RdRp and 0.1 mM ATP, and 0.5 mM CTP/UTP/GTP in the RdRp buffer (50 mM Hepes, 10 mM NaCl in pH 7.5) in 60 μl and incubated for 45 min at 37°C. NGO solution was prepared at 2.5 μg/ml in distilled water right before use from a stock solution (1 mg/ml). After the incubation, NGO was added to each sample at a final concentration of 1.5 μg/ml. Reactants were transferred to the microplate. The eventual RdRp activity was measured by monitoring the corresponding fluorescence intensity change at excitation (Ex)/emission (Em) = 650 nm/670 nm with a Synergy MX fluorometer (BioTek, UK).
3
Ce6 Cellular Uptake Quantification
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A total of 1×105 cells per well (2-ml cell suspension of SW480, HCT116, LoVo, and DLD1) were seeded in 6-well plates. The medium was then replaced with 2 ml of fresh medium, containing 5 µM Ce6, and incubated for 1–24 h. The solutions were removed, and the cells were rinsed with PBS (1 ml). Afterward, the cells were harvested and centrifuged at 1,500 rpm for 5 min. Pellets were subsequently washed with PBS (1 ml) and centrifuged again. The fluorescence of the supernatants was measured with a Synergy MX fluorometer (BioTek) with excitation and emission wavelengths of 500 and 670 nm, respectively. A calibration curve was then used to calculate the concentrations of Ce6 per 10,000 cells.
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