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MB Sample ID: SA022543
Local Sample ID: | LBMCFCDM48R3N |
Subject ID: | SU000463 |
Subject Type: | Cells |
Subject Species: | Homo sapiens |
Taxonomy ID: | 9606 |
Cell Biosource Or Supplier: | American Type Culture Collection |
Cell Strain Details: | Breast Cancer Cell lines (BT474, MCF-7, MDA-MB-231, MDA-MB-468) |
Cell Primary Immortalized: | Immortalized |
Species Group: | Human |
Select appropriate tab below to view additional metadata details:
Subject:
Subject ID: | SU000463 |
Subject Type: | Cells |
Subject Species: | Homo sapiens |
Taxonomy ID: | 9606 |
Cell Biosource Or Supplier: | American Type Culture Collection |
Cell Strain Details: | Breast Cancer Cell lines (BT474, MCF-7, MDA-MB-231, MDA-MB-468) |
Cell Primary Immortalized: | Immortalized |
Species Group: | Human |
Factors:
Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
---|---|---|---|---|
LBMCFCDM48R3N | SA022543 | FL006431 | Media alone | Treatment |
Collection:
Collection ID: | CO000457 |
Collection Summary: | Cell samples were scraped off the dish in 4 mL of 50:50 acetonitrile:water and collected into 15 mL tubes prior to homogenization and extraction. |
Sample Type: | cells |
Storage Conditions: | -80°C |
Treatment:
Treatment ID: | TR000477 |
Treatment Summary: | Briefly, mycoplasma tested and certified BCa cell lines will be purchased from American Type Culture Collection (ATCC) and maintained under the manufacturer’s recommended conditions. Cells were grown for 48 h in the presence or absence of chemotherapeutic agent paclitaxel (10 nM). |
Sample Preparation:
Sampleprep ID: | SP000470 |
Sampleprep Summary: | Added 1 mL of an ice-cold chloroform to the tubes and 10 ceramic beads. Tubes were vortexed for 30 seconds on a multi-tube vortexer at 5000 rpm three times to homogenize and centrifuged samples at 4 °C in a swinging bucket centrifuge for 60 min at 3,700 rpm. Majority of aqueous (top) layer was carefully transferred to 5 mL cryotubes. Majority of lipid (bottom) layer was carefully transferred to 7 mL glass vials. Remaining protein layer and residual aqueous & lipid layers were transferred into a 2 mL Lo-Bind tubes. Original tubes were rinsed with 600 µL of 2:1 chloroform:methanol solution and transferred to the 2 mL tubes. Centrifuged tubes at 15,000 rpm at 4 °C for 20 min and transferred remaining aqueous & lipid fractions into respective tubes. For each study sample, an 800 µL aliquot of the aqueous fractions was transferred to labeled 2.0 mL Lo-Bind tubes. Analytical pooled QC samples were generated by transferring an additional 75 µL aliquot from all study samples into a 10 mL tube. The total pooled sample was vortexed and 800 µL aliquots were transferred to 2.0 mL tubes labeled Pool. All tubes were frozen for 1 hr and lyophilized to dryness overnight. Samples were reconstituted by adding 700 µL of 90:10 D2O:Chenomx ISTD master mix to each, vortexed for 2 mins and centrifuged at 16,000 rcf for 4 min. A 600 µL aliquot of the supernatant was transferred into 5 mm NMR tubes (Bruker-Biospin, Switzerland), and kept on ice until data acquisition. |
Processing Method: | lyophilization |
Extraction Method: | Acetonitrile:water and chloroform |
Sample Resuspension: | NMR master mix |
Cell Type: | breast cancer |
Analysis:
MB Sample ID: | SA022543 |
Analysis ID: | AN000693 |
Laboratory Name: | David H. Murdock Research Institute. |
Analysis Type: | NMR |
Acquisition Date: | 7/29/14 and 8/13/14 |
Software Version: | TopSpin 3.2 |
Operator Name: | Jason Winnike |
Detector Type: | NMR |
Num Factors: | 2 |
NMR:
NMR ID: | NM000077 |
Analysis ID: | AN000693 |
Instrument Name: | Bruker Avance III 600 MHz |
Instrument Type: | FT-NMR |
NMR Experiment Type: | 1D 1H |
Field Frequency Lock: | Deuterium |
Standard Concentration: | 0.5 mM DSS |
Spectrometer Frequency: | 600 MHz |
NMR Probe: | 5 mm ATMA Cryoprobe |
NMR Solvent: | D2O |
NMR Tube Size: | 5 mm |
Shimming Method: | Topshim |
Pulse Sequence: | noesyprid |
Water Suppression: | yes |
Pulse Width: | 9.28 us |
Power Level: | 39.811 W |
Receiver Gain: | 50.8 |
Offset Frequency: | 2817.50 Hz |
Presaturation Power Level: | 0.000025119 W |
Chemical Shift Ref Cpd: | DSS |
Temperature: | 298.1 K |
Number Of Scans: | 256 |
Dummy Scans: | 4 |
Acquisition Time: | 2.482 s |
Relaxation Delay: | 2 s |
Spectral Width: | 11.0011 ppm, 6602.113 Hz |
Num Data Points Acquired: | 32768 |
Real Data Points: | 16384 |
Line Broadening: | 0.5 Hz |
Zero Filling: | yes |
Apodization: | Lorentzian |
Baseline Correction Method: | Polynomial |
Chemical Shift Ref Std: | DSS |
Binned Increment: | 0.04 |
Binned Data Excluded Range: | water (4.66 - 4.86 ppm); imidazole (7.15-7.26 ppm) |