Experiments were performed on 500 MHz, 750 MHz and 800 MHz Varian spectrometers. Resonances were assigned unambiguously using samples with site-specific low-enrichment 15N or 15N, 13C labeling and through-bond correlations at natural abundance. Standard 2D NMR experimental spectra including NOESY, TOCSY and COSY, were collected at 25 °C to obtain the complete proton resonance assignments. The NMR experiments for samples in water solution were performed with Watergate or Jump-and-Return water suppression techniques. The variable temperature spectra were recorded on 500 MHz Varian spectrometer from 5 °C to 50 °C. All spectra were processed by the program NMRPipe. Spectral assignments were also carried out and supported by COSY, TOCSY and NOESY spectra. Peak assignments were achieved using the software Sparky.
Varian spectrometer
Manufactured by Agilent Technologies
Sourced in United States, Germany
The Varian spectrometer is a laboratory instrument used for spectroscopic analysis. It is designed to measure the absorption or emission of electromagnetic radiation by samples, allowing for the identification and quantification of chemical compounds.
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Lab products found in correlation
Avance 400 spectrometer, by Bruker (2 mentions)
Dmf d7, by Merck Group (1 mentions)
Maxis 4g q tof mass spectrometer, by Bruker (1 mentions)
Onenmr probe, by Agilent Technologies (1 mentions)
Mestrenova software, by Mestrelab Research (1 mentions)
Nicolet is5 spectrometer, by Thermo Fisher Scientific (1 mentions)
1260 infinity isocratic pump, by Agilent Technologies (1 mentions)
Silica gel 60, by Merck Group (1 mentions)
Triple resonance cryoprobe, by Agilent Technologies (1 mentions)
17 protocols using varian spectrometer
1
NMR Characterization of Biomolecular Samples
2
Polymer Characterization by NMR and MALDI-TOF
3
NMR Characterization of Pyrazole Complexes
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An appropriate amount of the complexes 1a ,b and 2a ,b was dissolved in 200 µL of DMF-d7 and 300 µL of D2O was added (1 mM final concentration). 1H NMR spectra (Varian spectrometer (Varian Inc.; Palo Alto, CA, USA), 400 MHz, 298 K) of these solutions were recorded at various time points (0–48 h), and the solutions were stored at r.t. in the dark between the individual experiments. The spectra were referenced to the residual signal of water (4.79 ppm). Note: DMF-d7 ensured the solubility of the complexes, as their solubility in water is not sufficient for 1H NMR. Deuterated solvents DMF-d7 and D2O were supplied by Merck/Sigma-Aldrich (Prague, Czech Republic). Free pyrazoles were studied by 1H NMR under the same experimental conditions (for comparative purposes).
4
Characterization of Novel Organic Compounds
5
NMR Spectral Characterization Parameters
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Spectra were recorded in a 400 MHz Varian spectrometer, using a spectrometer frequency of 400.14 MHz with a OneNMR Probe and a ProTune System (Agilent). Spectral size range covered from −2 to 10 ppm. Receiver gain was fixed to 34. Also, 512 scans were used with a relaxation delay of 5 seconds. Spectral size contained 65 k data points, and the acquired size was made of 32 k complex data points.
6
Quantitative Metabolite Analysis of Cell Cultures
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1H-NMR spectra were acquired using a 600 MHz Varian spectrometer (Varian, Inc., Palo Alto, CA, USA) equipped with a 3-mm indirect detection probe, as previously described [25 (link)]. A total of 180 µL of medium of each condition (n = 15) was analyzed. Samples were diluted with a sodium fumarate and deuterated water solution (final concentration of 2 mM in 225 µL), which was used as internal reference (6.50 ppm) to quantify the following metabolites present in the medium (multiplet, ppm): lactate (doublet, 1.33), acetate (singlet, 1.90), and malate (double doublet, 2.37). Spectra were manually phased and baseline corrected. Chosen metabolites peaks were integrated using the NUTS-Pro NMR software (Acorn NMR, Inc, Fremont, CA, USA). Results are expressed as nmol/106 spermatozoa.
7
Polysaccharide and Oligosaccharide NMR Analysis
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The polysaccharide and purified oligosaccharides were analyzed by NMR spectroscopy. The samples were exchanged twice with 99.9% D2O by lyophilization and then dissolved in 0.6 mL of 99.96% D2O. Spectra were recorded on a 500 MHz VARIAN spectrometer (Varian Inc., Palo Alto, CA, USA) operating at 50 °C for the polymer and 25 °C for oligosaccharides solutions. Two-dimensional (2D) experiments were performed using standard VARIAN pulse sequences and pulsed field gradients for coherence selection when appropriate. Standard parameters were used for 2D NMR experiments. Chemical shifts are expressed in ppm using acetone as internal reference (2.225 ppm for 1H and 31.07 ppm for 13C). NMR spectra were processed using MestreNova software (Mestrelab Research, S.L., Santiago de Compostela, Spain).
8
NMR Characterization of Diluted Nanotubes
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NMR spectra were
recorded on a 500
MHz Varian spectrometer equipped with triple-resonance 3.2 mm T3 and
BioMAS probes. The MAS rates ranged between ∼8–11 kHz ± 5 Hz for the different experiments, and the sample
temperature was regulated at ∼10 °C. Spectra were processed
using NMRPipe52 (link) and analyzed using nmrglue.53 (link) The intramolecular 13C–13C and 13C–15N distances were
determined for the diluted nanotube sample using the rotational resonance
width33 (link) and z-filtered TEDOR experiments,32 (link) respectively, while the intermolecular 13C–15N distances were probed using the mixed
nanotube sample and band-selective TEDOR,32 (link) as described in detail in theSupporting Information .
recorded on a 500
MHz Varian spectrometer equipped with triple-resonance 3.2 mm T3 and
BioMAS probes. The MAS rates ranged between ∼8–11 kHz ± 5 Hz for the different experiments, and the sample
temperature was regulated at ∼10 °C. Spectra were processed
using NMRPipe52 (link) and analyzed using nmrglue.53 (link) The intramolecular 13C–13C and 13C–15N distances were
determined for the diluted nanotube sample using the rotational resonance
width33 (link) and z-filtered TEDOR experiments,32 (link) respectively, while the intermolecular 13C–15N distances were probed using the mixed
nanotube sample and band-selective TEDOR,32 (link) as described in detail in the
9
NMR Structural Determination of IFITM3
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IFITM3 proteinfor NMR experiments were expressed and purified as described before, except that the final NMR samples were prepared in 50 mM phosphate buffer containing 10% D2O, with a concentration of 1.0 mM. A series of TROSY-based multi-dimensional NMR experiments were performed on 500 MHz Varian spectrometer, 700 MHz Varian spectrometer and 600 MHz Bruker spectrometer. 1H/15N labeled IFITM3 protein was used for 2D TROSY-HSQC and NOESY-HSQC experiments. The 13C/15N labeled sample and 2H/13C/15N labeled sample in DPC detergent micelles were both applied for 3D HNCA, HN(CO)CA, HNCO, HN(CA)CO, CBCANH, and CBCA(CO)NH spectra collecting at pH 7.0 and 35 °C. Besides, to assist assignment, 2D TROSY experiments were performed using 15N-Leucine, 15N-Isoleucine, 15N-Valine, 15N-Methionine and 15N-Phenylalanine selectively labeled IFITM3 samples. Details of the experimental parameters were described previously. All the spectra were analyzed using NMRPipe and NMRView. Backbone resonance assignment was then performed. Dihedral angle constraints were predicted using Talos + program, based on the obtained chemical shifts of 13Cα, 13Cβ, 13CO, 1Hα, 15N, 1HN. Finally, the resulting dihedral angle and NOE restrains were applied in structure calculation using Xplor-NIH program.
10
NMR Spectroscopy and GPC Analysis of Polymers
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1H NMR spectra were recorded
using a 500 MHz Varian spectrometer. The CDCl3 singlet
at 7.26 ppm was selected as the reference standard. Spectral features
are tabulated in the following order: chemical shift (ppm); multiplicity
(s-singlet, d-doublet, t-triplet, m-complex multiple); number of protons;
position of protons. For the synthesis of (co)polymers, monomer conversion
was determined by 1H NMR analysis. Molecular weight and
molecular weight distribution of the (co)polymers were determined
by gel permeation chromatography (GPC). An Agilent GPC was equipped
with a 1260 Infinity Isocratic Pump and a RI detector. Two Agilent
PLgel mixed-C and mixed-D columns were used with DMF containing 0.1
mol % LiBr at 50 °C at a flow rate of 1.0 mL/min. Linear poly(methyl
methacrylate) standards from Fluka were used for calibration. Aliquots
of the polymer samples were dissolved in DMF/LiBr. The clear solutions
were filtered using a 0.40 μm PTFE filter to remove any DMF-insoluble
species. A drop of anisole was added as a flow rate marker. Fourier
transform infrared spectroscopy (FT-IR) was recorded on a Nicolet
iS5 spectrometer (Thermo Scientific).
using a 500 MHz Varian spectrometer. The CDCl3 singlet
at 7.26 ppm was selected as the reference standard. Spectral features
are tabulated in the following order: chemical shift (ppm); multiplicity
(s-singlet, d-doublet, t-triplet, m-complex multiple); number of protons;
position of protons. For the synthesis of (co)polymers, monomer conversion
was determined by 1H NMR analysis. Molecular weight and
molecular weight distribution of the (co)polymers were determined
by gel permeation chromatography (GPC). An Agilent GPC was equipped
with a 1260 Infinity Isocratic Pump and a RI detector. Two Agilent
PLgel mixed-C and mixed-D columns were used with DMF containing 0.1
mol % LiBr at 50 °C at a flow rate of 1.0 mL/min. Linear poly(methyl
methacrylate) standards from Fluka were used for calibration. Aliquots
of the polymer samples were dissolved in DMF/LiBr. The clear solutions
were filtered using a 0.40 μm PTFE filter to remove any DMF-insoluble
species. A drop of anisole was added as a flow rate marker. Fourier
transform infrared spectroscopy (FT-IR) was recorded on a Nicolet
iS5 spectrometer (Thermo Scientific).
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