The binding of AMDX with TTR was characterized by measuring the fluorescent spectra. AMDX compound solution 4 µM was prepared with and without 5 µM of aggregated TTR in triplicates. The maximum absorbance, emission, and excitation wavelength of AMDX-9101 with and without aggregated TTR was measured on a Shimadzu RF-5301PC spectrophotometer with the LabSolutions RF software (Shimadzu, Kyoto, Japan). The fold increase was calculated by measuring the relative fluorescent unit of AMDX + TTR/AMDX. The binding affinity (Kd) of AMDX-9101 and TTR was obtained by measuring the emission of increasing AMDX-9101 (0–10 µM) with a constant TTR concentration (5 µM) using the previously measured maximum excitation of 440 nm. The Kd was determined by plotting the relative fluorescent unit at maximum emission (λmax em) with TTR versus probe concentration.
Labsolutions rf software
Manufactured by Shimadzu
Sourced in Japan
LabSolutions RF software is a data processing and analysis tool designed for use with Shimadzu's RF series of fluorescence spectrophotometers. The software enables users to acquire, process, and analyze fluorescence data generated by the instruments.
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2 protocols using labsolutions rf software
1
Characterizing AMDX-TTR Binding Affinity
2
Characterization of Magnetic Nanoparticles in Cell Culture
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The MFNP characterization was performed using cell culture under similar conditions, at a concentration of 10 µg Fe/mL dispersed in DMEN/F-12 medium, supplemented with 10% FBS, and 1% antibiotic-antimycotic solution.
The MFNP optical characteristics were measured using an RF-6000 spectrofluorophotometer (Shimadzu, Kyoto, Japan) and LabSolutions RF software (Shimadzu, Kyoto, Japan). MFNP were dispersed in supplemented medium enclosed in a quartz cuvette with 1 cm of the optical path. The 2D excitation spectrum graphics were obtained in the wavelength between 400 to 850 nm and emissions from 450 to 870 nm.
The hydrodynamic size of MFNP was determined using the dynamic light scattering (DLS) technique with the Zetasizer Ultra system (Malvern, Worcestershire, UK). The polydispersion curve of hydrodynamic size was obtained at an angle of 173°, with the number of averages set at 30 and acquisition time of 3 s in a fixed position at 37 °C, with 120 s of thermic equilibrium. The mean diameter and standard deviation were calculated by adjusting the experimental data to a log-normal distribution function.
The colloidal stability analysis of hydrodynamic diameter over time was performed with MFNP dispersed in the supplemented medium. The polydispersion curves were recorded every hour for 18 h at 37 °C.
The MFNP optical characteristics were measured using an RF-6000 spectrofluorophotometer (Shimadzu, Kyoto, Japan) and LabSolutions RF software (Shimadzu, Kyoto, Japan). MFNP were dispersed in supplemented medium enclosed in a quartz cuvette with 1 cm of the optical path. The 2D excitation spectrum graphics were obtained in the wavelength between 400 to 850 nm and emissions from 450 to 870 nm.
The hydrodynamic size of MFNP was determined using the dynamic light scattering (DLS) technique with the Zetasizer Ultra system (Malvern, Worcestershire, UK). The polydispersion curve of hydrodynamic size was obtained at an angle of 173°, with the number of averages set at 30 and acquisition time of 3 s in a fixed position at 37 °C, with 120 s of thermic equilibrium. The mean diameter and standard deviation were calculated by adjusting the experimental data to a log-normal distribution function.
The colloidal stability analysis of hydrodynamic diameter over time was performed with MFNP dispersed in the supplemented medium. The polydispersion curves were recorded every hour for 18 h at 37 °C.
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