Optical rotations were measured using a P-1020 polarimeter (JASCO). UV spectrua were obtained with a DU 640 spectrophotometer (Beckman). ECD spectra were acquired on a JASCO J-815-150S CD spectrometer. IR spectra were obtained via a Nicolet-Nexus-470 spectrometer. NMR spectra were recorded on an Agilent DD2 NMR spectrometer (500 MHz). ESIMS and HRESIMS spectra were obtained by a Q-TOF (Micromass) and a LTQ Orbitrap XL (Thermo Scientific) spectrometer, respectively. Single-crystal analysis were performed on a Gemini A Ultra system using Cu Kα radiation (Aglient Technologies). A 1525 separation module (Waters) equipped with a C18 (Kromasil, 5 μm, 10 × 250 mm) column was used for semi-preparative HPLC. ODS (Unicorn; 45–60 μm), Sephadex LH-20 (Amersham Biosciences), and silica gel (200–300 mesh; Qing Dao Hai Yang Chemical Group Co.) were applied for column chromatography. TLC (G60, F-254; Yan Tai Zi Fu Chemical Group Co.) was used in the compounds detection.
J 815 150s cd spectrometer
Manufactured by Jasco
The J-815-150S CD spectrometer is a laboratory instrument designed for circular dichroism (CD) spectroscopy. It measures the difference in absorption of left-handed and right-handed circularly polarized light by a sample, providing information about the structural and conformational properties of biomolecules and other chiral materials.
Automatically generated - may contain errors
Lab products found in correlation
1525 separation module, by Waters Corporation (1 mentions)
Dd2 nmr spectrometer, by Agilent Technologies (1 mentions)
Ltq orbitrap xl, by Thermo Fisher Scientific (1 mentions)
Sephadex lh 20, by Cytiva (1 mentions)
P 1020 polarimeter, by Jasco (1 mentions)
Du 640 spectrophotometer, by Beckman Coulter (1 mentions)
Superdex 200 10 300 gl column, by Cytiva (1 mentions)
Amicon ultra 0.5 centrifugal filter device, by Merck Group (1 mentions)
Suprasil quartz cuvette, by Hellma (1 mentions)
3 protocols using j 815 150s cd spectrometer
1
Spectroscopic Analysis of Compounds
2
Circular Dichroism Spectroscopy Analysis
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Samples for circular dichroism (CD) were prepared by buffer exchanging the purified protein into CD buffer (0.1% LDAO, 20 mM NaPi pH 6.5, 100 mM NaF) using a Superdex 200 10/300 GL column (Cytiva). The eluent was concentrated using 30 kDa Amicon Ultra-0.5 centrifugal filter devices (MilliporeSigma) to a concentration of 1 mg/mL. Protein samples were loaded onto a 0.1 mm pathlength Suprasil quartz cuvette (Hellma) for CD measurement.CD spectra were collected using a JASCO J-815-150S CD spectrometer with the following parameters: wavelength range of 260-175 nm, speed of 100 nm/min, data pitch of 0.5 nm, D.I.T of 2 sec, bandwidth of 1 nm, and 7 accumulations for each sample. The secondary structure was estimated using JASCO’s multivariate secondary structure estimation program. Thermal melt curves were generated by using the spectrophotometer with a JASCO PTC-423 Peltier controller accessory. CD spectra were collected at different temperatures starting from 20 °C and up to 90 °C at 5 °C increments.
3
Secondary Structure Analysis of NSD Proteins
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The Circular Dichroism (CD) spectroscopy measurement was performed to investigate the secondary structure composition of the NSD proteins. Protein samples were mixed with 200 mM KCl buffer and CD spectra were recorded from 240 nm to 190 nm at 20 °C for 60 cycles using a 0.1 mm cuvette in a J-815-150S CD Spectrometer (Jasco). As background signal, dialysis buffer II (20 mM Tris/HCl pH 7.2, 100 mM KCl, 0.5 mM DTT, 60% glycerol) was measured under the same conditions.
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