Absorptions spectra were recorded at RT on a 4000 UV–Vis-spectrophotometer (Varian). Fluorescence spectra were recorded at 77 K on a Fluorolog 3.22 spectrofluorimeter (Jobin Yvon-Spex). All fluorescence spectra were measured at an OD of < 0.05 cm−1 at the Qy maximum. The circular-dichroism (CD) spectra were recorded on a Chirascan-Plus spectropolarimeter (Applied Photophysics). When necessary, the samples were diluted with a 20 mM MOPS (pH 7.5) or 20 mM MES (pH 5.5) buffer containing 10 mM NaCl. Plots were generated in OriginPro 2020 (OriginLab).
Fluorolog 3.22 spectrofluorimeter
Manufactured by SPEX
The Fluorolog 3.22 spectrofluorimeter is a laboratory instrument designed for the measurement of fluorescence spectra. It is capable of performing steady-state and time-resolved fluorescence measurements.
Automatically generated - may contain errors
Lab products found in correlation
Chirascan plus spectropolarimeter, by Applied Photophysics (2 mentions)
4000 uv vis spectrophotometer, by Agilent Technologies (1 mentions)
Originpro 2020, by OriginLab (1 mentions)
Nitrogen cryostat, by Oxford Instruments (1 mentions)
Cary 4000 spectrophotometer, by Agilent Technologies (1 mentions)
Chirascan cd spectrophotometer, by Applied Photophysics (1 mentions)
3 protocols using fluorolog 3.22 spectrofluorimeter
1
Spectroscopic Characterization of Biomolecules
2
Biophysical Characterization of Pigments
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Room temperature absorption spectra were recorded with a Cary 4000 spectrophotometer (Varian). The CD spectra were measured using a Chirascan CD Spectrophotometer (AppliedPhotophysics). The fluorescence emission spectra were recorded using a Fluorolog 3.22 spectrofluorimeter (Jobin Yvon-Spex). The excitation wavelengths were 440nm, 475nm and 500nm and emission was detected in the 600–800nm range. Excitation and emission bandwidths were set to 3 nm. All fluorescence spectra were measured at OD 0.05 at the maximum of the Qy absorption band. Room temperature measurements were performed in 0.5M sucrose, 20mM Hepes (pH 7.5), 0.06% β-DM buffer. For 77K measurements a liquid nitrogen cooled device was used (Nitrogen Cryostat, Oxford Instruments). The samples were in 70% glycerol (w/v), 20 mM Hepes (pH 7.5), 0.06% β-DM buffer.
3
Spectroscopic Characterization of Photosynthetic Pigments
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The absorption spectra at room temperature were recorded on a Varian Cary 4000 UV-vis-spectrophotometer with a 0.5 nm step from 350 to 750 nm. Fluorescence spectra were recorded at room temperature with an OD of 0.05/cm at the Qy maximum on a Fluorolog 3.22 spectrofluorimeter (Jobin Yvon-Spex). Fluorescence spectra at 77 K were measured on the same set-up using a cold finger filled with liquid nitrogen. Circular-dichroism (CD) spectra were recorded with a 1 nm step from 350 to 750 nm at 20 °C on a Chirascan-Plus spectropolarimeter (Applied Photophysics). When necessary the samples were diluted with a sucrose buffer (0.5 M sucrose, 10 mM Hepes, 0.03% α-DDM, pH 7.5).
Pigment analysis was performed as described in Xu et al. [30] with slight modification in the HPLC program. The amount of buffer B was linearly increased from 0% to 100% in 9.2 min without stop. SDS-PAGE electrophoresis was performed with 4-12% gradient precast gels from Invitrogen.
Pigment analysis was performed as described in Xu et al. [30] with slight modification in the HPLC program. The amount of buffer B was linearly increased from 0% to 100% in 9.2 min without stop. SDS-PAGE electrophoresis was performed with 4-12% gradient precast gels from Invitrogen.
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