This study was performed in vitro, using fluorescence measurements from a Hitachi-F3010 spectrofluorometer (Tokyo, Japan). Albumins (codes A-0281 and A-8763) and sulpiride (S2191000) were purchased from Sigma-Aldrich Co., St. Louis, MO, USA.
F3010 spectrofluorometer
Manufactured by Hitachi
Sourced in Japan
The F3010 spectrofluorometer is a laboratory instrument designed for the measurement of fluorescence spectra. It features a high-performance monochromator and a sensitive detector to analyze the emission and excitation characteristics of fluorescent samples.
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2 protocols using f3010 spectrofluorometer
1
Albumin-Sulpiride Fluorescence Assay
2
Quantitative ROS Measurement in U87MG Cells
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U87MG cells were seeded at a density of 1×10 5 cells per well in 24-well plates and incubated for 24 h at 37 °C. Cells were then treated with CPT formulations equivalent to 20 ng/mL CPT for 24 h, washed twice with PBS and fixed with 4% paraformaldehyde for 10 min. After washing two times with PBS, cells were incubated with DCFDA (10 µM) for 30 min in the dark. Cells were finally washed with PBS and observed using a fluorescence microscope (Nikon, Japan).
For quantitative estimation of ROS, cells were harvested using 0.25% trypsin-EDTA and washed with PBS. Cell counts were adjusted for all formulations. Cells were incubated with DCFDA (10 µM) for 30 min in the dark and fluorescence intensity was measured at excitation and emission wavelengths of 495 and 520 nm, respectively, using a Hitachi F-3010 spectrofluorometer.
For quantitative estimation of ROS, cells were harvested using 0.25% trypsin-EDTA and washed with PBS. Cell counts were adjusted for all formulations. Cells were incubated with DCFDA (10 µM) for 30 min in the dark and fluorescence intensity was measured at excitation and emission wavelengths of 495 and 520 nm, respectively, using a Hitachi F-3010 spectrofluorometer.
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