NMR spectra of 4.4 mg/mL of rat HspB2/B3 in 90% H2O/10% D2O, 20 mM phosphate buffer (pH 7.5) and 100 mM NaCl were acquired at 25 °C and a 1H frequency of 600 MHz on a Varian Unity Inova 600 NMR spectrometer. The two-dimensional 1H–1H TOCSY spectrum had a spin lock mixing time of 80 ms and the 1H–1H nuclear Overhauser effect spectroscopy spectrum had a mixing time of 100 ms.
Unity inova 600 nmr spectrometer
Manufactured by Agilent Technologies
Sourced in United States
The Unity Inova 600 NMR spectrometer is a nuclear magnetic resonance (NMR) instrument designed for analytical purposes. It provides high-resolution NMR spectroscopy capabilities for the identification and characterization of chemical compounds.
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5 protocols using unity inova 600 nmr spectrometer
1
NMR Spectroscopy of Rat HspB2/B3
2
SCFA Analysis After Allo-HSCT
3
NMR Analysis of Serum Samples
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Before nuclear magnetic resonance (NMR) experimentation, serum samples were thawed at room temperature for no longer than 20 min and 400 μL aliquots were combined with 100 μL of saline (0.9% NaCl in 10% D2O/90% H2O) and centrifuged at 12,000g for 5 min. A 400‐μL aliquot of this solution was put into a 5‐mm NMR tube for NMR analysis.
1H NMR analysis was performed on a Varian Unity INOVA 600 NMR spectrometer (Palo Alto, CA) operating at a frequency of 599.93 Hz. All experiments were run with 128 scans and a recycle time delay of 2.1 s to ensure a steady state of recovered magnetization and 64k time domain points. A Carr‐Purcell‐Meiboom‐Gill sequence was used to record the one‐dimensional spectra with a relaxation delay of 100 ms.
1H NMR analysis was performed on a Varian Unity INOVA 600 NMR spectrometer (Palo Alto, CA) operating at a frequency of 599.93 Hz. All experiments were run with 128 scans and a recycle time delay of 2.1 s to ensure a steady state of recovered magnetization and 64k time domain points. A Carr‐Purcell‐Meiboom‐Gill sequence was used to record the one‐dimensional spectra with a relaxation delay of 100 ms.
4
NMR Analysis of Serum Samples
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For NMR analysis, the samples were thawed at room temperature. Serum samples were prepared by mixing 200 μL serum with 400 μL saline buffer solution (45 mM NaH2PO4, 45 mM K2HPO4, 0.9% saline in a 20% v/v D2O, 80% v/v H2O solvent, pH 7.4). The serum-buffer mixture was kept at room temperature for 10 min then centrifuged at 10,000 rpm/min for 10 min. The clarified supernatant (550 μL) was transferred into a 5 mm NMR tube for spectroscopic analysis. The serum 1H NMR spectrum of each sample was recorded using the Carr-Purcell-Meiboom-Gill pulse sequence (RD − 90° − [τ − 180° − τ]n − AQ) on a Varian Unity Inova600 NMR spectrometer at a proton frequency of 599.95 MHz. For each sample, 128 scans were collected with 32,768 data points over a 10,000 Hz spectral width, resulting in a 1.64 s acquisition time and a 2 s relaxation delay at 298 K. The water signal was suppressed using a presaturation sequence on the water-signal frequency during the relaxation delay. For assignment purposes, selected samples were subjected to two-dimensional NMR, including 1H- 1H homonuclear correlation spectroscopy (COSY), total correlation spectroscopy (TOCSY), and J-resolved spectroscopy (J-Res).
5
NMR Characterization of Synthetic DNA
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The DNAs utilized in NMR studies were synthesized using Dr Oligo 48 DNA synthesizer (Biolytic Lab Performance Inc, USA), dialyzed successively three times against ddH2O, lyophilized and dissolved in 500 μl of three differ buffers. Buffer 1 and 2 are composed of 50 mM CAPS-NaOH pH 10.5, 1.0 M (NH4)2SO4 and 100 mM Li2SO4, and 50 mM CAPS-NaOH pH 9.2, 100 mM (NH4)2SO4 and 100 mM Li2SO4, respectively. Buffer 3 is composed of 80 mM NaH2PO4/Na3PO4 pH 6.8, 100 mM (NH4)2SO4 and 100 mM Li2SO4. A total of 10% D2O is included in all NMR buffers. The concentrations of each NMR sample were typically about 1.5–2.0 mM. The one-dimensional 1H NMR spectra, with a spectral width 16 ppm and scanning number 2K, were performed at 20°C on a Varian Unity Inova 600 NMR spectrometer equipped with a triple resonances cryoprobe and pulsed field gradients.
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