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Drx avance 700 mhz spectrometer

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The DRX Avance 700 MHz spectrometer is a nuclear magnetic resonance (NMR) instrument designed for high-resolution spectroscopy. It operates at a frequency of 700 MHz, providing enhanced sensitivity and resolution for the analysis of chemical and biological samples.

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Proteinase k, by Serva Electrophoresis (2 mentions) Dnase, by Serva Electrophoresis (2 mentions)

3 protocols using drx avance 700 mhz spectrometer

1

Isolation and Purification of Bacterial O-Antigens

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The LPS was isolated applying the hot phenol-water method55 , followed by dialysis against distilled water until the phenol scent was gone. Then samples were treated with DNase (1mg/100 mg LPS) plus RNase (2 mg/100 mg LPS) at 37°C for 2 h, followed by Proteinase K treatment (1 mg/100 mg LPS) at 60°C for 1 h [all enzymes from Serva, Germany]. Subsequently, samples were dialyzed again for 2 more days, then freeze dried. Such LPS samples were then hydrolyzed with 1% aqueous acetic acid (100°C, 90 min) and ultra-centrifuged for 16 h at 4°C and 150,000 g. Resulting supernatants (the O-antigens) were dissolved in water and freeze-dried. For further purification, the crude O-antigen samples were chromatographed on TSK HW-40 eluted with pyridine/acetic acid/water (10/4/1000, by vol.), then lyophilized. On these samples, 1D and 2 D (COSY, TOCSY, HSQC, HMBC) 1H- and 13C-NMR spectra were recorded with a Bruker DRX Avance 700 MHz spectrometer (1H: 700.75 MHz; 13C: 176.2 MHz) as described56 (link).
Diard M., Bakkeren E., Lentsch V., Rocker A., Bekele N.A., Hoces D., Aslani S., Arnoldini M., Böhi F., Schumann-Moor K., Adamcik J., Piccoli L., Lanzavecchia A., Stadtmueller B.M., Donohue N., van der Woude M.W., Hockenberry A., Viollier P.H., Falquet L., Wüthrich D., Bonfiglio F., Loverdo C., Egli A., Zandomeneghi G., Mezzenga R., Holst O., Meier B.H., Hardt W.D, & Slack E. (2021). A rationally designed oral vaccine induces Immunoglobulin A in the murine gut that directs the evolution of attenuated Salmonella variants. Nature microbiology, 6(7), 830-841.
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2

Isolation and Purification of Bacterial O-Antigens

Check if the same lab product or an alternative is used in the 5 most similar protocols
The LPS was isolated applying the hot phenol-water method55 , followed by dialysis against distilled water until the phenol scent was gone. Then samples were treated with DNase (1mg/100 mg LPS) plus RNase (2 mg/100 mg LPS) at 37°C for 2 h, followed by Proteinase K treatment (1 mg/100 mg LPS) at 60°C for 1 h [all enzymes from Serva, Germany]. Subsequently, samples were dialyzed again for 2 more days, then freeze dried. Such LPS samples were then hydrolyzed with 1% aqueous acetic acid (100°C, 90 min) and ultra-centrifuged for 16 h at 4°C and 150,000 g. Resulting supernatants (the O-antigens) were dissolved in water and freeze-dried. For further purification, the crude O-antigen samples were chromatographed on TSK HW-40 eluted with pyridine/acetic acid/water (10/4/1000, by vol.), then lyophilized. On these samples, 1D and 2 D (COSY, TOCSY, HSQC, HMBC) 1H- and 13C-NMR spectra were recorded with a Bruker DRX Avance 700 MHz spectrometer (1H: 700.75 MHz; 13C: 176.2 MHz) as described56 (link).
Diard M., Bakkeren E., Lentsch V., Rocker A., Bekele N.A., Hoces D., Aslani S., Arnoldini M., Böhi F., Schumann-Moor K., Adamcik J., Piccoli L., Lanzavecchia A., Stadtmueller B.M., Donohue N., van der Woude M.W., Hockenberry A., Viollier P.H., Falquet L., Wüthrich D., Bonfiglio F., Loverdo C., Egli A., Zandomeneghi G., Mezzenga R., Holst O., Meier B.H., Hardt W.D, & Slack E. (2021). A rationally designed oral vaccine induces Immunoglobulin A in the murine gut that directs the evolution of attenuated Salmonella variants. Nature microbiology, 6(7), 830-841.
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3

Structural Elucidation of Lipid A by NMR

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Homo- and heteronuclear 1D (1H, 13C) and 2D NMR experiments, i.e., correlation spectroscopy (1H,1H-COSY), double-quantum filtered phase sensitive correlation spectroscopy (DQF-COSY), total correlation spectroscopy (1H,1H-TOCSY), rotating frame nuclear Overhauser effect spectroscopy (1H,1H-ROESY), heteronuclear single quantum coherence-distortionless enhancement by polarization transfer spectroscopy (1H,13C-HSQC-DEPT), and heteronuclear multiple bond correlation (1H,13C-HMBC) were recorded on solutions of lipid A in DMSO-2H6/C2H3Cl (1:1, v:v) at 27 °C with a Bruker DRX Avance 700 MHz spectrometer that was equipped with a 5 mm CPQCI multinuclear-inverse cryo probe head with a z gradient and Bruker software. The used frequencies were 700.75 MHz for 1H NMR and 176.2 MHz for 13C NMR. The NMR spectra of de-acylated lipid A sample, which were obtained from mild hydrazinolysis and hot KOH treatment, were recorded on a solution of 2H2O. All 1H,13C spectra were calibrated to internal acetone (δH 2.225, δC 31.45).
Komaniecka I., Susniak K., Choma A., Heine H, & Holst O. (2020). The Lipid A from the Lipopolysaccharide of the Phototrophic Bacterium Rhodomicrobium vannielii ATCC 17100 Revisited. International Journal of Molecular Sciences, 22(1), 258.
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