Prior to NMR analysis, the dried cerebellum samples (NWT = 5 and NKlf10 KO = 5) were reconstituted in 220 μL of deuterated buffer containing TSP (3-trimethylsilylpropionic acid) at 145 μmol·L−1 and transferred to conventional 3 mm NMR tubes (CortecNet). As previously described, the extraction protocol of metabolites from cerebellum was adapted from the Folch type two-step procedure described by Wu et al. [27 (link)]. 1H-NMR spectra were obtained with a Bruker DRX-600 AVANCE-III HD spectrometer (Bruker SADIS, Wissembourg, France), operating at 14 T, with a TCI cryoprobe. Standard water suppressed 1H-NMR spectra were acquired at 298 K using a “noesypr1d” pulse sequence with relaxation delay of 20 s. Spectra were processed using Topspin version 3.2 software (Bruker Daltonik, Karlsruhe, Germany). As previously described [28 (link)], 1H-NMR spectra were automatically reduced to ASCII files using AMIX Software package (Analysis of MIXture, version 3.9.14, Bruker Biospin, Karlsruhe, Germany). Spectral intensities were scaled to the total spectral intensity, and the resulting data table was analyzed by multivariate and univariate statistical analyses.
Analysis of mixture
Manufactured by Bruker
Sourced in Germany
The Analysis of MIXture is a laboratory equipment product designed for the analysis of complex mixtures. It provides advanced analytical capabilities to researchers and scientists. The core function of this product is to enable the identification and quantification of individual components within a mixture sample.
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2 protocols using analysis of mixture
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Metabolite Profiling of Cerebellum Samples
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Metabolite Profiling of Cerebellum Samples
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Prior to NMR analysis, the dried cerebellum samples (NWT = 5 and NKlf10 KO = 5) were reconstituted in 220 μL of deuterated buffer containing TSP (3-trimethylsilylpropionic acid) at 145 μmol·L−1 and transferred to conventional 3 mm NMR tubes (CortecNet). As previously described, the extraction protocol of metabolites from cerebellum was adapted from the Folch type two-step procedure described by Wu et al. [27 (link)]. 1H-NMR spectra were obtained with a Bruker DRX-600 AVANCE-III HD spectrometer (Bruker SADIS, Wissembourg, France), operating at 14 T, with a TCI cryoprobe. Standard water suppressed 1H-NMR spectra were acquired at 298 K using a “noesypr1d” pulse sequence with relaxation delay of 20 s. Spectra were processed using Topspin version 3.2 software (Bruker Daltonik, Karlsruhe, Germany). As previously described [28 (link)], 1H-NMR spectra were automatically reduced to ASCII files using AMIX Software package (Analysis of MIXture, version 3.9.14, Bruker Biospin, Karlsruhe, Germany). Spectral intensities were scaled to the total spectral intensity, and the resulting data table was analyzed by multivariate and univariate statistical analyses.
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