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Cpmgpr1d

Manufactured by Bruker
Sourced in Germany

The Cpmgpr1d is a nuclear magnetic resonance (NMR) spectroscopy instrument designed for performing one-dimensional (1D) experiments. It is capable of acquiring high-resolution NMR spectra of liquid samples.

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8 protocols using cpmgpr1d

1

NMR-based Lipoprotein Subclass Analysis

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Serum levels of apoB-containing lipoproteins were measured on a Bruker 600 MHz Avance Neo NMR spectrometer using the Bruker IVDr lipoprotein subclass analysis protocol, as described previously [7 (link),48 (link)]. Briefly, serum samples were thawed, and 330 µL of each sample was mixed with 330 µL of Bruker serum buffer (Bruker, Rheinstetten, Germany). The samples were mixed gently and 600 µL of the mixed sample was transferred into a 5 mm SampleJet rack tube (Bruker, Rheinstetten, Germany). Proton spectra were obtained at a constant temperature of 310 K using a standard Nuclear Overhauser Effect Spectroscopy (NOESY) pulse sequence (Bruker: noesygppr1d), a Carr–Purcell–Meiboom–Gill (CPMG) pulse sequence with presaturation during the relaxation delay (Bruker: cpmgpr1d) to achieve water suppression, and a standard 2D J-resolved (JRES) pulse sequence (Bruker: jresgpprqf). Data analysis was carried out using the Bruker IVDr Lipoprotein Subclass Analysis (B.I.LISA™, Bruker, Rheinstetten, Germany) method. Lipid contents of apoB-containing lipoprotein particles were calculated as ratios of serum levels of the respective lipid in apoB-containing lipoprotein (mg/dL) and apoB in apoB-containing lipoprotein (mg/dL).
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2

Lipoprotein Profiling by NMR Spectroscopy

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Blood serum lipoproteins were measured on a Bruker 600 MHz Avance Neo NMR spectrometer using the Bruker IVDr lipoprotein subclass analysis protocol. Serum samples were thawed, and 330 µL of each sample mixed with 330 µL of Bruker serum buffer (Bruker, Rheinstetten, Germany). The samples were mixed gently, and 600 µL of the mixed sample were transferred into a 5 mm SampleJet rack tube (Bruker). Proton spectra were obtained at a constant temperature of 310 K using a standard nuclear Overhauser effect spectroscopy (NOESY) pulse sequence (Bruker: noesygppr1d), a Carr–Purcedll–Meiboom–Gill (CPMG) pulse sequence with presaturation during the relaxation delay (Bruker: cpmgpr1d) to achieve water suppression, and a standard 2D J-resolved (JRES) pulse sequence (Bruker: jresgpprqf). Data analysis was carried out using the Bruker IVDr LIpoprotein Subclass Analysis (B.I.LISATM, Bruker, Rheinstetten, Germany) method.
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3

NMR Analysis of HDL Subclasses

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Serum levels of total HDL-C, HDL-TG, HDL-PL, HDL-apoA-I, and HDL-apoAII, as well as of their 4 size/density subclasses (HDL1: 1.063–1.100 kg/L; HDL2: 1.100–1.112 kg/L; HDL3: 1.112–1.125 kg/L; HDL4: 1.125–1.210 kg/L), were measured on a Bruker 600 MHz Avance Neo NMR spectrometer using the Bruker IVDr lipoprotein subclass analysis protocol, as described [17 (link),60 (link)]. Briefly, serum samples were thawed, and 330 µL of each sample was mixed with 330 µL of Bruker serum buffer (Bruker, Rheinstetten, Germany). The samples were mixed gently and 600 µL of the mixed sample was transferred into a 5 mm SampleJet rack tube (Bruker). Proton spectra were obtained at a constant temperature of 310 K using a standard Nuclear Overhauser Effect Spectroscopy (NOESY) pulse sequence (Bruker: noesygppr1d), a Carr–Purcell–Meiboom–Gill (CPMG) pulse sequence with presaturation during the relaxation delay (Bruker: cpmgpr1d) to achieve water suppression, and a standard 2D J-resolved (JRES) pulse sequence (Bruker: jresgpprqf). Data analysis was carried out using the Bruker IVDr LIpoprotein Subclass Analysis (B.I.LISATM, Bruker Biospin, Rheinstetten, Germany).
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4

NMR-based Metabolomic Analysis of Plasma

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Plasma samples were prepared for metabolomic analysis based on nuclear magnetic resonance (NMR) by mixing 3:1 (v/v) of samples and citrate buffer (pH 4.4) using an automated mixer on board. The analysis was performed on an automated high-throughput NMR platform with an Agilent spectrometer 400 MHz (9.4 T), a 4 mm indirect detection probe, and a fixed flow cell (Agilent Technologies, Santa Clara, CA, USA). Data were acquired and processed using Topspin 3.6 (Bruker Biospin), and experiments were performed under automation by the IconNMR program (Bruker Biospin) using the standard pulse sequence, cpmgpr1d (Bruker Biospin).
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5

High-Throughput Serum NMR Profiling

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The 1H NMR spectra of all the samples were acquired on 800 MHz NMR spectrometer (BrukerAvance-III) equipped with the cryoprobe at 300 Kelvin (K). On each serum sample, the 1D 1H transverse relaxation-edited CPMG (Carr–Purcell–Meiboom–Gill) NMR spectra were recorded using the standard Bruker’s pulse program library sequence (cpmgpr1d) with pre-saturation of the water peak through irradiating it continuously during the recycle delay (RD) of 5 sec. Each spectrum consisted of the accumulation of 128 scans and lasted for approximately 15 min. To remove broad signals from proteins and fats, a total spin–spin relaxation time of 60 ms (n=300 and 2τ=200μs) was applied. Each FID (free induction decay) was zero filled and Fourier-transformed to 64 K data points following manual phase and baseline-correction. A line broadening factor of 0.3 Hz and a sine–bell apodization function was applied to FIDs before Fourier Transformation (FT). The raw NMR data were processed using Bruker software Topspin-v2.1 (BrukerBioSpin GmbH, Silberstreifen 4 76287 Rheinstetten, Germany).
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6

NMR Analysis of Serum Lipoproteins

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Blood serum lipoproteins were analyzed on a Bruker 600 MHz Avance Neo NMR spectrometer using the Bruker IVDr lipoprotein subclass analysis protocol. Serum samples were thawed, and 330 µL of each sample mixed with 330 µL of Bruker serum buffer (Bruker, Rheinstetten, Germany). The samples were mixed gently, and 600 µL of the mixed sample were transferred into a 5 mm SampleJet rack tube (Bruker, Rheinstetten, Germany). Proton spectra were obtained at a constant temperature of 310 K using a standard nuclear Overhauser effect spectroscopy (NOESY) pulse sequence (Bruker: noesygppr1d), a Carr–Purcedll–Meiboom–Gill (CPMG) pulse sequence with presaturation during the relaxation delay (Bruker: cpmgpr1d) to achieve water suppression, and a standard 2D J-resolved (JRES) pulse sequence (Bruker: jresgpprqf) [9 (link)]. Data analysis was carried out using the Bruker IVDr Lipoprotein Subclass Analysis (B.I.LISATM) method.
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7

Probing Breast Cancer Tissue Metabolism

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Frozen human breast cancer tissue samples were cut to fit a leak-proof 30-μL disposable insert (mean sample weight: 8.8 mg) added 3 μL of phosphate-buffered saline (PBS) based on D2O with trimethylsilyl propionate (TSP, 1 mM) and sodium formate (1 mM). The insert was placed in a 4-mm diameter zirconium rotor (Bruker, Biospin GmbH, Germany). Spin-echo experiments (cpmgpr1D; Bruker, L4 = 136) were run with 2 ms delay between 180° pulses, TE of 273.5 ms, spectral width of 20 ppm (−5 to 15 ppm) and NS of 256 scans (17 (link)). To evaluate the effect of prolonged HR MAS MRS experimental time, data acquisition was repeated after 1.5 h. The sample was kept spinning (5000 Hz) within the magnet at 5°C in this time interval.
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8

High-Res NMR Metabolomics of Tissue

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A total of 270 tissue samples were analyzed by high resolution (HR) magic angle spinning (MAS) NMR. Details of NMR experiments, preprocessing and quantification of the tissue samples have been described previously. 21 Briefly, tissue samples (mean weight: 4.1 mg) were analysed at 5 °C on a Bruker Avance DRX600 spectrometer equipped with a 1 H/ 13 C MAS rotor. A spin-echo one dimensional experiment with presaturation (cpmgpr1d, Bruker BioSpin, Germany) was recorded for all samples, with effective echo time of 77 ms, a spectral width of 20 ppm (-5 to 15 ppm), and 256 scans.
Spectra were baseline corrected, peak aligned using the icoshift algorithm 31 , and normalized by PQN 34 after removal of lipid residuals. Quantified metabolites were normalized by PQN.
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