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MB Sample ID: SA011335
| Local Sample ID: | folch_low |
| Subject ID: | SU000266 |
| Subject Type: | Human cells |
| Subject Species: | Homo sapiens |
| Taxonomy ID: | 9606 |
| Cell Biosource Or Supplier: | ATCC - Supplier |
| Subject Comments: | P?+10 |
| Cell Primary Immortalized: | immortalized |
| Cell Passage Number: | P?+10 |
| Cell Counts: | P?+10 |
| Species Group: | Human |
Select appropriate tab below to view additional metadata details:
Subject:
| Subject ID: | SU000266 |
| Subject Type: | Human cells |
| Subject Species: | Homo sapiens |
| Taxonomy ID: | 9606 |
| Cell Biosource Or Supplier: | ATCC - Supplier |
| Subject Comments: | P?+10 |
| Cell Primary Immortalized: | immortalized |
| Cell Passage Number: | P?+10 |
| Cell Counts: | P?+10 |
| Species Group: | Human |
Factors:
| Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
|---|---|---|---|---|
| folch_low | SA011335 | FL002781 | Folch | Extraction Method |
Collection:
| Collection ID: | CO000257 |
| Collection Summary: | - |
| Sample Type: | Cells - Jurkat, Clone E6-1 |
| Collection Method: | Centrifugation |
| Collection Frequency: | 1200 rpm |
| Collection Duration: | 5 min |
| Volumeoramount Collected: | 10 mL of cell suspension |
| Storage Conditions: | -80°C |
| Tissue Cell Quantity Taken: | 1e6 cells |
Treatment:
| Treatment ID: | TR000277 |
| Cell Storage: | Liquid Nitrogen, 5% DMSO |
| Cell Growth Container: | 10 cm2 culture dish |
| Cell Media: | RPMI 1640 |
| Cell Envir Cond: | 95% humidity, 5% CO2 |
| Cell Harvesting: | centrifugation |
Sample Preparation:
| Sampleprep ID: | SP000271 |
| Sampleprep Summary: | Cells were extracted using either the Folch or Matyash method. The total volume of MeOH in each extraction method was varied (0.5 mL or 2.0 mL) while the ratio of MeOH to the other solvents was maintained. |
| Sampleprep Protocol Filename: | SOP_LipidExtractionProtocol.pdf |
| Processing Method: | cell lysis |
| Processing Storage Conditions: | on ice |
| Extraction Method: | Folch or Matyash lipid extraction method |
| Extract Concentration Dilution: | 1e6 cells were extracted with the solvent systems explained in the Sample Prep Summary |
| Sample Resuspension: | 100 μL of IPA |
| Sample Spiking: | internal standards: lysophosphatidylcholine (LPC 17:0 and LPC 19:0), phosphatidylcholine (PC 17:0/17:0 and PC 19:0/19:0), phosphatidylethanolamine (PE 15:0/15:0 and PE 17:0/17:0), phosphatidylserine (PS 14:0/14:0 and PS 17:0/17:0), and phosphatidylglycerol (PG 14:0/14:0 and PG 17:0/17:0), triacylglyceride (TG 15:0/15:0/15:0 and TG 17:0/17:0/17:0) |
| Cell Type: | Jurkat, Clone E6-1 |
Combined analysis:
| Analysis ID | AN000384 |
|---|---|
| Analysis type | MS |
| Chromatography type | Reversed phase |
| Chromatography system | Thermo Dionex Ultimate 3000 RS |
| Column | Supelco Analytical Titan C18 |
| MS Type | ESI |
| MS instrument type | Orbitrap |
| MS instrument name | Thermo Q Exactive Orbitrap |
| Ion Mode | POSITIVE |
| Units | Peak Area |
Chromatography:
| Chromatography ID: | CH000276 |
| Chromatography Summary: | Reversed Phase |
| Methods Filename: | m_lipid_pos_switch_100 |
| Instrument Name: | Thermo Dionex Ultimate 3000 RS |
| Column Name: | Supelco Analytical Titan C18 |
| Column Pressure: | 1241 bar (max) |
| Column Temperature: | 30 C |
| Flow Gradient: | 32% B at 0 min, 40% B at 1 min, a hold at 40% B until 1.5 min, 45% B at 4 min, 50% B at 5 min, 60% B at 8 min, 70% B at 11 min, and 80% B at 14 min |
| Flow Rate: | 0.5 mL/min |
| Injection Temperature: | 4 C |
| Sample Injection: | 2 μL |
| Solvent A: | 40% water/60% acetonitrile; 0.1% formic acid; 10 mM ammonium formate |
| Solvent B: | 90% isopropanol/8% acetonitrile/2% water; 0.1% formic acid; 10 mM ammonium formate |
| Analytical Time: | 21 min |
| Randomization Order: | 3,10,12,11,8,6,5,4,7,9,2,1 |
| Chromatography Type: | Reversed phase |
MS:
| MS ID: | MS000330 |
| Analysis ID: | AN000384 |
| Instrument Name: | Thermo Q Exactive Orbitrap |
| Instrument Type: | Orbitrap |
| MS Type: | ESI |
| Ion Mode: | POSITIVE |
| Capillary Temperature: | 320 |
| Dry Gas Flow: | 30 |
| Mass Accuracy: | <2 ppm |
| Spray Voltage: | 3.5 kV |
| Desolvation Gas Flow: | .raw |
| Scan Range Moverz: | 100-1500 |
| Skimmer Voltage: | 15 V |
| Acquisition Parameters File: | QE1_CZU_08_sequence |
