Fluorescence spectra were acquired with an FLSP920 spectrofluorometer (Edinburgh Instruments Ltd, Livingston, UK) and the chamber temperature was controlled by a circulator (Julabo Labortechnik GmbH, Seelbach, Germany) at 20 ± 1°C. Fluorescence was measured in a quartz cell with a path length of 1 cm. Ligand at the specified concentration was mixed with the DNA solution; the resulting solution was incubated for 30 min at room temperature and was then equilibrated for 5 min in the chamber before fluorescence determination. The Job's plot analysis was used to fluorescently evaluate the binding stoichiometry of ligand with DNA. The total concentration of ligand and DNA was kept at 2 μM and the concentration ratio was systematically changed. The SFA-20 stopped-flow accessory (TgK Scientific Ltd, UK) was used to fluorescently check the switching kinetics (excitation/emission: 440/490 nm).
Flsp920 spectrofluorometer
Manufactured by Edinburgh Instruments
Sourced in United Kingdom
The FLSP920 spectrofluorometer is a laboratory instrument designed for the measurement of fluorescence spectra. It features high-performance optics and a sensitive detector to enable accurate and reliable fluorescence analysis.
Automatically generated - may contain errors
4 protocols using flsp920 spectrofluorometer
1
Fluorescence Spectroscopy of Ligand-DNA Binding
2
Fluorescence Spectroscopy of Heme
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Fluorescence spectra were recorded with FLSP920 spectrofluorometer (Edinburgh Instruments, UK). Spectra from samples containing 1 μM heme in MeHCl were obtained by summation of four independent measurements. All fluorescence spectra were obtained with a 10 × 10 mm quartz cell and corrected for the wavelength-dependent sensitivity of the detection.
3
Steady-State Luminescence Characterization
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Steady state luminescence, excitation and lifetime measurements were performed using a FLSP920 spectrofluorometer (Edinburgh Instruments) equipped with double-monochromators, a continuous Xe-lamp of 450 W and a pulsed Xe-lamp of 60 W (µF920) for excitation, and a Hamamatsu R928 photomultiplier tube (PMT) for detection. All emission spectra were corrected for the system response. The excitation and emission spectra were normalized to the intensity of the 7 F 0 → 5 L 6 and 5 D 0 → 7 F 2 transitions, respectively, and an offset was applied for the sake of clarity.
4
Characterization of Synthetic Compounds
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All of the reagents and solvents were obtained commercially and used without any additional purification. The assessment of the progress of reactions and purity of the synthesized compounds was performed using TLC on MERCK Silica gel 60 F254 plates, with hexane/acetone (1:2) as an eluent, and visualized under UV light. Melting points were determined using a METTLER TOLEDO™ Melting Point System MP70 apparatus. The IR spectrum was recorded on a Thermo Scientific Nicolet iS50 Spectrometer (ATR accessory; no. of scans: 64; resolution: 4; data spacing: 0.482 cm−1). 1H-, 13C-, and 31P-NMR were captured using a Bruker Avance 500 MHz (Bruker Corporation, Billerica, MA, USA) in CDCl3 at room temperature. Solvent peaks were used as the internal reference. Chemical shift (δ) values are reported in ppm. Accurate high-resolution mass measurements were conducted using the Orbitrap Exploris 120 (Thermo Fisher Scientific, Waltham, MA, USA), operating in Full Scan mode at a resolution of 120,000. The FLSP920 spectrofluorometer (Edinburgh Instruments Ltd., Livingston, UK) captured fluorescence emission spectra within the visible range of 450–800 nm, while the absorption spectra were acquired using the UV-visible spectrophotometer SPECORD® 80 (Analytik Jena AG, Jena, Germany).
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