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Standard 5 mm nmr tube

Manufactured by Wilmad
Sourced in United States
About the product

The Standard 5 mm NMR tube is a precision-made glass vessel used for nuclear magnetic resonance (NMR) spectroscopy analysis. It is designed to hold liquid samples during NMR measurements, providing a consistent and reliable environment for data collection.

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12 protocols using «standard 5 mm nmr tube»

1

NMR Metabolic Profiling Protocol

All procedures were previously described in Duft12 (link). The sample preparation included the washing of the filters 3 kDa (Amicon Ultra) with H20 Milli-Q and centrifugation at 20,817g (14,000 rpm in an Eppendorf 5417R) for 10 min at 4 °C. Then, the samples were centrifugated in the same rotation and temperature for 45 min. The filtered solution (200 µl) was transferred to a standard 5 mm NMR tube (Wilmad), and added 60 μL of phosphate buffer (pH 7.4 for the pH standardization), containing 0.5 mM of TMSP for internal chemical shift reference and 340 μL of Milli-Q H2O, completing a total of 600 μL in the NMR tube.
The spectra acquisition was carried out by nuclear magnetic resonance spectroscopy (1H NMR) in a 600 MHz spectrometer (Varian Inova. Agilent Technologies. Santa Clara. CA), equipped with a triple cold probe. A total of 256 scans were collected with an acquisition time of 4 s and relaxation delay intervals between scans of 1.5 s. The temperature was maintained at 298 K (25 °C). After the acquisition, we realized the phase adjustment, baseline correction, spectral calibration, and quantification of metabolites, conducted by the software Chenomx NMR Suite 7.6 (Chenomx. Edmonton. AB. Canada)12 (link) (Fig. 5).

Representation of the 1H NMR spectrum region where the main metabolites were quantified. (A) Glutamine. (B) Glucose. (C) Pyruvate (D) 2-Oxoisocaproate and (E) 3-hydroxyisobutyrate.

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2

NMR Sample Preparation Protocol

2020
All standards for the development of the
database were purchased from Sigma–Aldrich (Prague, Czech Republic)
as well as the dimethyl-silapentane-sulfonate (DSS) used as an internal
standard. Deuterium oxide (99.96% D) was purchased from Eurisotop
(Saarbrücken, Germany). Syringes, syringe filters (PVDF membrane,
0.2 μm), and vials were purchased from Fisher Scientific (Pardubice,
Czech Republic). Standard 5 mm NMR tubes were purchased from Wilmad-LabGlass
(Warminster, PA, U.S.A.).
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3

Parahydrogen-Enhanced NMR Spectroscopy

2018
pH2 was prepared using commercially available setup with a conversion temperature of 38 K that provides ≈ 90 % enrichment of the para fraction (Bruker, BPHG 90). Experiments were carried out inside a 300 MHz NMR spectrometer (Bruker AVANCE). The reaction chamber for pH2 supply was described before38 (link). pH2 is delivered in a standard 5 mm NMR tube (Wilmad) via a thin 1/16” capillary. pH2 flow rate was 7 scm3/min with 1.7 bar overpressure with respect to an ambient pressure.
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4

NMR Spectroscopy for Hydrophilic Metabolite Analysis

2017
All NMR spectra were acquired at 293 K on a Varian 800 MHz NMR spectrometer (operating at 799.42 MHz for 1H) equipped with a Z-axis-gradient 5 mm HCN probe. The hydrophilic metabolites were reconstituted in 600 μl of D2O containing 0.05 mM 4,4-dimethyl-4-silapentane-1-sulfonic acid (DSS) for chemical shift reference and internal concentration standard and 0.2% sodium azide (w/v) as bacteriostatic agent to prevent biodegradation. Approximately 550 μl of the prepared sample was loaded into a standard 5 mm NMR tube (Wilmad-LabGlass, Buena, NJ). One-dimensional 1H NMR spectra were acquired from each sample using the standard Varian PRESAT pulse sequence with a single excitation and 1.5 s low-power pre-saturation at the water peak position to suppress the residual water signal. A total of 6 k transients were accumulated to ensure a high quality 1H spectrum was obtained with sufficient signal-to-noise ratio for metabolites with concentration as low as approximately 0.5 μM in the NMR tube. For metabolite signal assignment and confirmation purposes, two-dimensional (2D) NMR spectra, including 1H–1H correlation spectroscopy (COSY) and 1H J-resolved spectroscopy (JRES), were acquired at 293 K for selected samples.
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5

NMR Kinetic Studies of Isocitrate Lyase

NMR experiments were conducted at a 1H frequency of 500 MHz using a Bruker Avance III HD spectrometer equipped with a BBFO probe. Experiments were conducted at 300 K. Standard 5 mm NMR tubes (Wilmad) using a sample volume of 500 μL were used in all experiments. The pulse tip-angle calibration using the single-pulse nutation method (Bruker pulsecal routine) was undertaken for each sample.60 (link) All measurements were performed in triplicate.
Time course experiments were monitored by standard Bruker 1H experiments with water suppression by excitation sculpting. Unless otherwise stated, the number of transients was 16, and the relaxation delay was 2 seconds. The lag time between addition of enzyme and the end of the first experiment was usually 4 minutes. Initial rates were calculated by linear fitting using Excel 2013 (Microsoft) for data points up to 20% turnover. Kinetic parameters were obtained using the Hanes plot. Linear fitting was done using Excel 2013 (Microsoft). All NMR samples contained 190 nM ICL1, 1 mM dl-isocitrate and 5 mM MgCl2 buffered with 50 mM Tris-D11 (pH 7.5) in 90% H2O and 10% D2O. For kinetic parameter measurements, the isocitrate concentrations ranged from 50 μM to 750 μM. For single concentration inhibition assay, 100 μM inhibitors was used. For IC50 measurements, varying concentrations of inhibitors were used. IC50 values were obtained by SigmaPlot 13 and fitted with the Sigmoid, 3 parameter model.
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