Biochemical properties of different-age extracted collagens were evaluated after solubilization at 2 mg/ml in 0.018 M acetic acid (v/v). Firstly, electrophoretic properties were estimated by 5% sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) after denaturation through heating at 90°C for 2 min. Gels were stained with Coomassie Brillant Blue R250 for revealing characteristic bands of type I collagen (α1 and α2 chains). Secondly, the fluorescence specific to AGEs was measured using a Shimadzu RF-5000 spectrofluorometer (Shimadzu, France) at 380 nm excitation and 440 nm emission. AGE-modified bovine serum albumin (BSA) was used to create a standard curve.
Rf 5000 spectrofluorometer
Manufactured by Shimadzu
Sourced in France
The RF-5000 spectrofluorometer is a laboratory instrument designed to measure the fluorescence properties of samples. It is capable of exciting samples with a specific wavelength of light and detecting the resulting fluorescence emission. The core function of the RF-5000 is to provide quantitative and qualitative analysis of fluorescent compounds in a variety of applications.
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Lab products found in correlation
5 protocols using rf 5000 spectrofluorometer
1
Biochemical Characterization of Collagens
2
Measuring Lipid Membrane Fluidity with DPH
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A thin film of 1,6-diphenyl-1,3,5-hexatriene (DPH) dissolved in chloroform was deposited along the inner walls of a glass vial and dried with a current of N 2 followed by desiccation. The liposome solution, warmed at 62℃, was added to the film so that the final DPH concentration was 300 times less than the lipid concentration of the liposome 22) . The material was vortex mixed and stored in dark overnight at 25℃. The fluorescence intensities were measured at different temperatures within the range of 25 to 45℃ using RF-5000 spectrofluorometer (Shimadzu Co., Ltd) . The emitted light (wavelength＝450 nm) polarized parallel and perpendicular to the excitation radiation (wavelength＝350 nm) was used for these measurements. From the obtained fluorescence intensities, the fluorescence anisotropy (r) values were calculated. Fluorescence anisotropy (r) for highly turbid samples were corrected for light scattering effects using a method described elsewhere 9, 23) .
3
Fluorescence Anisotropy of Lipid Membranes
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A thin film of 1,6-diphenyl-1,3,5-hexatriene dissolved in chloroform was deposited along the inner walls of a glass vial. The solvent was removed by passing a stream of N 2 gas followed by vacuum desiccation. The liposome solution, warmed at 62℃, was added to the film so that the final DPH concentration was 300 times less than the lipid concentration of the liposome 11) . The material was vortex mixed and kept overnight in the dark at 25℃. The samples of each composition were labeled in the same manner. The fluorescence intensities were measured at different temperatures in the range 25-45℃ using RF-5000 spectrofluorometer (Shimadzu Co., Ltd) . The emitted light polarized parallel and perpendicular to the excitation radiation was used for these measurements. The excitation and emission wavelengths were 350 nm and 450 nm, respectively. From the obtained fluorescence intensities, the fluorescence anisotropy (r) values were calculated. Labeling of the membrane with the ANS (1-anilinonaphthalene-8-sulfonic acid) probe was carried out by a similar method using its ethanolic solution (0.1 mM) . The excitation and emission wavelengths chosen in this case were 380 and 465 nm, respectively. r for highly turbid samples were corrected for light scattering effects using a method described elsewhere 12) .
4
Spectroscopic Characterization of Palladium(II) Complex
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Human serum albumin (HSA) was purchased from Sigma-Aldrich. 2-Hydroxy-1-naphtaldehhyde and allylamine were obtained from Merck Co. and used without further purification. All salts used for buffer preparation were analytical grade and dissolved in double distilled water. All of the solutions were used freshly after preparation. The FT-IR spectra were recorded on a JASCO, FT/IR-6300 spectrometer (4000-400 cm -1 ) in KBr pellets. 1 H and 13 C NMR spectra were recorded on a Bruker Avance 400 spectrometer using DMSO as a solvent for the Palladium(II) complex. The elemental analysis was performed on Leco, CHNS-932 and Perkin-Elmer 7300 DV elemental analyzers. The UV-vis spectra were recorded on a JASCO V-670 spectrophotometer.
Fluorescence measurements were carried out on Shimadzu RF-5000 spectrofluorometer at room temperature. The circular dichroism (CD) spectra were recorded using Aviv spectropolarimeter model 215 (Proterion Corp., USA) at 25 °C. The scan speed was 20 nm min -1 . The CD spectra were recorded in the far-UV region (190-260 nm) using 0.1 cm path length cell.
Fluorescence measurements were carried out on Shimadzu RF-5000 spectrofluorometer at room temperature. The circular dichroism (CD) spectra were recorded using Aviv spectropolarimeter model 215 (Proterion Corp., USA) at 25 °C. The scan speed was 20 nm min -1 . The CD spectra were recorded in the far-UV region (190-260 nm) using 0.1 cm path length cell.
5
Spectroscopic Characterization of Chiral Compound
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All required materials were purchased from Sigma-Aldrich except 2-Hydroxy-1-naphtaldehhyde and (R/S)-1-aminopropan-2-ol that were obtained from Merck Co. The FT-IR spectrum was recorded on a JASCO, FT/IR-6300 spectrometer (4000-400 cm -1 ) in KBr pellets. The elemental analysis was performed on Leco, CHNS-932 and Perkin-Elmer 7300 DV elemental analyzers. The UV-Vis spectra were recorded on a JASCO V-670 spectrophotometer. Fluorescence measurements were carried out on Shimadzu RF-5000 spectrofluorometer at room temperature.
The circular dichroism (CD) spectra were recorded using Aviv spectropolarimeter model 215 (Proterion Corp., USA) at 25 °C.
The circular dichroism (CD) spectra were recorded using Aviv spectropolarimeter model 215 (Proterion Corp., USA) at 25 °C.
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