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MB Sample ID: SA323578
Local Sample ID: | S_C0h_EA_13_1_2 |
Subject ID: | SU003095 |
Subject Type: | Cultured cells |
Subject Species: | Homo sapiens |
Taxonomy ID: | 9606 |
Genotype Strain: | MDA-MB-231 cells |
Gender: | Not applicable |
Cell Biosource Or Supplier: | ATCC (Manassas, VA, USA); ATCC HTB-26 |
Cell Strain Details: | Epithelial breast cancer cells; absence of estrogen and progesterone receptors, HER2 overexpression |
Cell Passage Number: | Inferior to 10 |
Cell Counts: | 5 M |
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Sample Preparation:
Sampleprep ID: | SP003101 |
Sampleprep Summary: | The cellular polar extracts were extracted using a biphasic extraction method of methanol/chloroform/water. Basically, cell pellets were resuspended in 650 µL of 80% (v/v) methanol-miliQ water solution, transferred to microcentrifuge tubes with 150 mg of glass beads, and vortexed for 5 min. Subsequently, 260 µL of 100% chloroform and 260 µL of 100% chloroform plus 220 µL MiliQ water were added to samples, which were vortexed for 5 min between solvents addition. The samples were kept at − 20 °C for 10 min and centrifuged. The aqueous phase of the resulting extract was collected into a new tube, vacuum-dried and stored at − 80 °C until the NMR analysis. All samples and reagents were kept in ice during the extraction procedure. Before NMR analysis, the dry aqueous extracts were suspended in 650 µL of 100 mM sodium phosphate buffer (pH 7.4, in D2O containing 0.25% 3-(trimethylsilyl)propionic-2,2,3,3-D4 acid sodium salt (TSP-D4) for chemical shift referencing) and transferred into 5 mm NMR tubes. |
Processing Storage Conditions: | On ice |
Extraction Method: | Biphasic extraction method of methanol/chloroform/water |
Extract Storage: | -80℃ |
Sample Resuspension: | Dry aqueous extracts were suspended in 650 µL of 100 mM sodium phosphate buffer (pH 7.4, in D2O containing 0.25% 3-(trimethylsilyl)propionic-2,2,3,3-D4 acid sodium salt (TSP-D4) for chemical shift referencing) |