NMR spectra were acquired at 4°C on a Bruker Avance III 500-MHz spectrometer, equipped with a 5-mm triple-resonance TCI cryogenic probe head. Spectra were processed using Topspin (Bruker Biospin Ltd.) and analyzed with CCPN (Collaborative Computing Project for NMR) analysis. The NOESY spectrum used a NOESY mixing time of 100 ms. Samples were prepared at a concentration of 430 μM (NOESY) or 100 μM (HSQC titrations of S100B or Aβ42), in 50 mM Hepes (pH 7.3), 5 mM DTT, 10% D2O, with or without 10 mM CaCl2. The chemical shift assignments for S100B under these conditions have been deposited in the BioMagResBank under the accession number 27199.
Tci cryogenic probe head
Manufactured by Bruker
The TCI cryogenic probe head is a specialized component used in nuclear magnetic resonance (NMR) spectroscopy. It is designed to operate at cryogenic temperatures, enhancing the sensitivity and resolution of NMR measurements. The TCI cryogenic probe head supports various NMR experimental techniques and is compatible with Bruker's NMR spectrometer systems.
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2 protocols using tci cryogenic probe head
1
NMR Characterization of S100B and Aβ42
2
NMR Analysis of Lactonized α2,3-Sialyllactose
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All NMR data was recorded on a Bruker AVANCE II spectrometer equipped with a 14.1 T magnet and a 5 mm TCI cryogenic probe head and a z-gradient system. The samples were manually injected into disposable 5 mm SampleJet NMR tubes and sealed with a closed cap. A Bruker SampleJet system was used for sample insertion, removal and temporary storage. In the SampleJet the samples were kept at a temperature of 6 °C. The temperature for the NMR measurements was 300 K, which was carefully calibrated using a fresh methanol-d4 sample. TSP was used as chemical shift reference. All sample were measured in an 1D NOESY 1H NMR experiment with 25 Hz water presaturation and a relaxation delay of 4 s. The reference α2,3-sialyllactose was measured by accumulating 4 scans, for the lactone sample 256 scans were collected. The lactonized α2,3-sialyllactose was analyzed directly after preparation, as well as after 16 h of storage at 300 K. The raw NMR data was processed and analyzed in Bruker TopSpin 3.0.
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