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MB Sample ID: SA354084
| Local Sample ID: | 210324_Rescue_Gust_compounds_2h_n1_Ti41_Cl_1uM_T50 |
| Subject ID: | SU003380 |
| Subject Type: | Cultured cells |
| Subject Species: | Homo sapiens |
| Taxonomy ID: | 9606 |
Select appropriate tab below to view additional metadata details:
Subject:
| Subject ID: | SU003380 |
| Subject Type: | Cultured cells |
| Subject Species: | Homo sapiens |
| Taxonomy ID: | 9606 |
Factors:
| Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
|---|---|---|---|---|
| 210324_Rescue_Gust_compounds_2h_n1_Ti41_Cl_1uM_T50 | SA354084 | FL041287 | Breast cancer cells | Sample source |
| 210324_Rescue_Gust_compounds_2h_n1_Ti41_Cl_1uM_T50 | SA354084 | FL041287 | 1 µM Comp. 3 | Treatment |
| 210324_Rescue_Gust_compounds_2h_n1_Ti41_Cl_1uM_T50 | SA354084 | FL041287 | 2 h | time point |
Collection:
| Collection ID: | CO003373 |
| Collection Summary: | Cultured cells were washed, trypsinized, counted and flash-frozen in liquid N2 and stored at -80°C. |
| Sample Type: | Breast cancer cells |
| Storage Conditions: | -80℃ |
Treatment:
| Treatment ID: | TR003389 |
| Treatment Summary: | Treatment of breast cancer cells (MDA-MB-231 cells) for the analysis of PE and PC: Human MDA-MB-231 breast cancer cells were treated with vehicle (DMSO) or RSL3 (1 or 10 µM) with or without ferrostatin-1 (3 µM) for 2 h, 4 h, 6 h, or 24 h or with SCs (1, 3, and 10 µM) for 2 h at 37°C and 5% CO2. Cells were harvested, washed with PBS pH 7.4, snap-frozen, and stored at -80°C. |
Sample Preparation:
| Sampleprep ID: | SP003387 |
| Sampleprep Summary: | Phospholipids were extracted from cell pellets by successive addition of PBS pH 7.4, methanol, chloroform, and saline to a final ratio of 14:34:35:17. Evaporation of the organic layer yielded a lipid film that was dissolved in methanol and subjected to UPLC-MS/MS. |
| Extract Storage: | -80℃ |
Combined analysis:
| Analysis ID | AN005344 |
|---|---|
| Analysis type | MS |
| Chromatography type | Reversed phase |
| Chromatography system | Waters Acquity H-Class |
| Column | Waters ACQUITY UPLC BEH C8 (100 x 2.1mm,1.7um) |
| MS Type | ESI |
| MS instrument type | QTRAP |
| MS instrument name | ABI Sciex 6500+ |
| Ion Mode | NEGATIVE |
| Units | relative intensities |
Chromatography:
| Chromatography ID: | CH004046 |
| Chromatography Summary: | Chromatographic separation of phospholipids was carried out on an Acquity BEH C8 column (1.7 μm, 130 Å, 2.1×100 mm, Waters, Milford, MA) using an ExionLC UHPLC system. |
| Instrument Name: | Waters Acquity H-Class |
| Column Name: | Waters ACQUITY UPLC BEH C8 (100 x 2.1mm,1.7um) |
| Column Temperature: | 45°C |
| Flow Gradient: | The gradient was ramped from 75 to 85% B over 5 min and further increased to 100% B within 2 min, followed by isocratic elution for another 2 min. |
| Flow Rate: | 0.75 mL/min |
| Solvent A: | 90% Water, 10% Acetonitrile; 2 mM ammonium acetate |
| Solvent B: | 5% Water, 95% Acetonitrile; 2 mM ammonium acetate |
| Chromatography Type: | Reversed phase |
MS:
| MS ID: | MS005074 |
| Analysis ID: | AN005344 |
| Instrument Name: | ABI Sciex 6500+ |
| Instrument Type: | QTRAP |
| MS Type: | ESI |
| MS Comments: | Targeted MRM with pre-optimized settings and subsequent automated integration of selected signals using Analyst 1.6.3 or Analyst 1.7.1 (Sciex). Relative intensities (indicating the proportion of lipids) were obtained by summing all signals analyzed within the subgroup (e.g., PE) and expressing the individual signals of lipid species or lipid subfractions as a percentage of this sum (= 100%). |
| Ion Mode: | NEGATIVE |
