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MB Sample ID: SA007479
Local Sample ID: | C33 |
Subject ID: | SU000156 |
Subject Type: | Human |
Subject Species: | Homo sapiens |
Taxonomy ID: | 9606 |
Species Group: | Human |
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Sample Preparation:
Sampleprep ID: | SP000153 |
Sampleprep Summary: | Serum samples were analyzed following filtration through a 3-kD nominal cut-off membrane (Amicon Ultra 0.5 mL centrifugal filters, Millipore Corp, Billerica, MA) to remove proteins, lipids, lipoproteins, and protein-bound forms of certain molecules. Each serum sample was transferred to a prewashed filter and centrifuged at 13,800 g for 60 minutes as previously described (Gregory, Park et al. 2013). The filters were further washed with deuterium oxide (D2O) and the filtrate was mixed with the filtered serum in a 1:1 ratio. A reference buffer solution containing 5 mmol/L disodium-2,2-dimethyl 2-silapentane-5-sulphonate (DSS) and 10 mmol/L imidazole in D2O was added to the samples in a 1:9 ratio. The D2O provided a field-frequency lock, whereas DSS was used as the chemical shift reference and imidazole as pH indicator for the NMR spectra. A total of 600 µL mixture was transferred to a 5 mm NMR tube (Sigma Aldrich, St. Louis, MO). |