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MB Sample ID: SA235401
Local Sample ID: | FContA-1-PE |
Subject ID: | SU002435 |
Subject Type: | Cultured cells |
Subject Species: | Homo sapiens |
Taxonomy ID: | 9606/10090 |
Select appropriate tab below to view additional metadata details:
Subject:
Subject ID: | SU002435 |
Subject Type: | Cultured cells |
Subject Species: | Homo sapiens |
Taxonomy ID: | 9606/10090 |
Factors:
Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
---|---|---|---|---|
FContA-1-PE | SA235401 | FL029198 | Control1 | Genotype |
Collection:
Collection ID: | CO002428 |
Collection Summary: | Fibroblasts and MEFs were cultured under 5% CO2 at 37°C in Dulbecco's modified medium (DMEM; Invitrogen, Carlsbad, CA) supplemented with 10% fetal bovine serum. For the extraction of phospholipids, fibroblasts were seeded in 6-well plate and cultured overnight, followed by replacement of fresh culture medium. Cells were harvested at 48 hours after the medium change as described below. |
Sample Type: | Cultured fibroblasts |
Treatment:
Treatment ID: | TR002447 |
Treatment Summary: | No treatment |
Sample Preparation:
Sampleprep ID: | SP002441 |
Sampleprep Summary: | Cells were detached by incubation with trypsin and suspended in PBS. Protein concentration was determined by the bicinchonic acid method (Thermo Fisher Scientific, Rockford, IL). Total lipids were extracted from 50 μg of cellular protein by the Bligh and Dyer method. Cells were dissolved in methanol/chloroform/water at 2:1:0.8 v/v/v and then 50 pmol of 1,2-adidodecanoyl-sn-glycero-3-phosphocholine (Avanti Polar Lipids, Alabaster, AL) and 1,2-didodecanoyl-sn-glycero-3-phosphoethanolamine (Avanti Polar Lipids) were added as internal standards. After incubation for 5 min at room temperature, 1 ml each of water and chloroform were added and the samples were then centrifuged at 2,000 rpm for 5 min in Himac CF-16RX (Hitachi Koki, Tokyo, Japan) to collect the lower organic phase. To re-extract the lipids from the water phase, 1 ml chloroform was added. The combined organic phase was evaporated under a nitrogen stream and the extracted lipids were dissolved in methanol. |
Combined analysis:
Analysis ID | AN003830 | AN003831 |
---|---|---|
Analysis type | MS | MS |
Chromatography type | Reversed phase | Reversed phase |
Chromatography system | Waters Acquity | Waters Acquity |
Column | Waters Aquity BEH C18 (150 x 1.0mm,1.7um) | Waters Aquity BEH C18 (150 x 1.0mm,1.7um) |
MS Type | ESI | ESI |
MS instrument type | Triple quadrupole | Triple quadrupole |
MS instrument name | ABI Sciex 4000 QTrap | ABI Sciex 4000 QTrap |
Ion Mode | POSITIVE | POSITIVE |
Units | The ratio to total amount of each phospholipid species | The ratio to total amount of each phospholipid species |
Chromatography:
Chromatography ID: | CH002835 |
Chromatography Summary: | PE and pPE detection method |
Instrument Name: | Waters Acquity |
Column Name: | Waters Aquity BEH C18 (150 x 1.0mm,1.7um) |
Chromatography Type: | Reversed phase |
Chromatography ID: | CH002836 |
Chromatography Summary: | PC and aPC detection method |
Instrument Name: | Waters Acquity |
Column Name: | Waters Aquity BEH C18 (150 x 1.0mm,1.7um) |
Chromatography Type: | Reversed phase |
MS:
MS ID: | MS003572 |
Analysis ID: | AN003830 |
Instrument Name: | ABI Sciex 4000 QTrap |
Instrument Type: | Triple quadrupole |
MS Type: | ESI |
MS Comments: | MS were detected by neutral loss, precoursour ion scan, and MRM. Peaks were detected by quantification wizard on Ananlyst software (version 1.42). |
Ion Mode: | POSITIVE |
MS ID: | MS003573 |
Analysis ID: | AN003831 |
Instrument Name: | ABI Sciex 4000 QTrap |
Instrument Type: | Triple quadrupole |
MS Type: | ESI |
MS Comments: | MS were detected by neutral loss, precoursour ion scan, and MRM. Peaks were detected by quantification wizard on Ananlyst software (version 1.42). |
Ion Mode: | POSITIVE |