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MB Sample ID: SA210159
| Local Sample ID: | Terence_MS2_Quad_Aq |
| Subject ID: | SU002275 |
| Subject Type: | Mammal |
| Subject Species: | Mus musculus |
| Genotype Strain: | C57BL/6J |
| Age Or Age Range: | 13 weeks |
| Weight Or Weight Range: | (28.4±1.1 g (control SHAM mice) vs 26.4±2.1 g (control AVF mice) vs 23.6±2.1 (CKD SHAM mice) vs 24.7±1.3 (CKD AVF mice), N=8-10/group). |
| Gender: | Male |
| Animal Animal Supplier: | Jackson Labs (Stock # 000664) |
| Animal Housing: | Housed in a temperature of 22 oC |
| Animal Light Cycle: | 12-hour light/12-hour dark |
| Animal Feed: | Ad libitum (Casein control diet vs. adenine-supplemented diet to induce CKD) |
| Animal Water: | free access to food and water (3-5 animals per cage). |
| Animal Inclusion Criteria: | (3-5 animals per cage). |
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Sample Preparation:
| Sampleprep ID: | SP002281 |
| Sampleprep Summary: | Weights of frozen quadriceps specimens were weighed using a microbalance (Mettler-Toledo, Columbus, OH, USA). Next, a slightly modified FOLCH extraction was performed to extract aqueous and lipid phase metabolites. The aqueous phase was lyophilized overnight (Labconco Corporation, Kansas, MO, USA) and the lipid phase was dried by passing inert nitrogen gas. The resulting aqueous and lipid phase dry powders were stored at -80oC until analysis using nuclear magnetic resonance (NMR). The dry powder of aqueous phase samples was dissolved in 50 µL of phosphate buffer system (50 mM, pH 7.2) consisting of 0.5 mM D6-DSS, 2 mM EDTA and 0.2% NaN2. Lipid phase dry powders were dissolved in 70 µL of CDCl3 supplemented with 10 mM of pyrazine (as internal NMR standard). All samples were loaded into 1.5 mm optical density (O.D.) NMR tubes. |
| Sampleprep Protocol Filename: | AVF_NMR_Aqueous_Phase_Procedures.docx |
| Processing Method: | Lyophilization and Homogenization |
| Processing Storage Conditions: | -80℃ |
| Extraction Method: | Modified FOLCH extraction |
| Extract Storage: | -80℃ |
| Sample Resuspension: | In 50 microliter of 50 mM phosphate buffer (pH 7.2) with 2 mM EDTA, 0.5 mM DSS and 0.2% sodium azide for aqueous phase samples. |
| Sample Spiking: | 0.5 mM of DSS for aqueous phase samples |
