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MB Sample ID: SA124026
Local Sample ID: | Nov5_2018_020 |
Subject ID: | SU001525 |
Subject Type: | Cultured cells |
Subject Species: | Homo sapiens |
Taxonomy ID: | 9606 |
Gender: | Female |
Select appropriate tab below to view additional metadata details:
Subject:
Subject ID: | SU001525 |
Subject Type: | Cultured cells |
Subject Species: | Homo sapiens |
Taxonomy ID: | 9606 |
Gender: | Female |
Factors:
Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
---|---|---|---|---|
Nov5_2018_020 | SA124026 | FL015001 | Control | siRNA |
Nov5_2018_020 | SA124026 | FL015001 | Vehicle (methanol) | Treatment (8hrs) |
Collection:
Collection ID: | CO001520 |
Collection Summary: | BT-549 cells were transfected with ACSL1 or non-targeting siRNA 72 hours prior to incubation with 25 uM alpha eleostearic acid or vehicle (methanol) for 8 hours. After 8 hours, cells (~4-6 x 106 per replicate) were trypsinized, collected, washed once with phosphate-buffered saline, and frozen in liquid nitrogen. |
Sample Type: | Breast cancer cells |
Storage Conditions: | -80℃ |
Treatment:
Treatment ID: | TR001540 |
Treatment Summary: | Cells were treated in complete culture medium with 25uM alpha eleostearic acid or methanol (vehicle control) for eight hours |
Treatment Compound: | alpha eleostearic acid |
Sample Preparation:
Sampleprep ID: | SP001533 |
Sampleprep Summary: | Cells were re-suspended in 25 mM HEPES buffer (pH 7.4) containing 200 μM DTPA and sonicated. |
Combined analysis:
Analysis ID | AN002425 | AN002426 |
---|---|---|
Analysis type | MS | MS |
Chromatography type | Normal phase | Reversed phase |
Chromatography system | Thermo Dionex Ultimate 3000 | Thermo Dionex Ultimate 3000 |
Column | Phenomenex Luna Silica (150 x 2.0mm,3um) | Phenomenex Luna Silica (150 x 1.0mm,3um) |
MS Type | ESI | ESI |
MS instrument type | Orbitrap | Orbitrap |
MS instrument name | Thermo Fusion Orbitrap | Thermo Q Exactive HF hybrid Orbitrap |
Ion Mode | NEGATIVE | POSITIVE |
Units | pmol/mg | pmol/umol |
Chromatography:
Chromatography ID: | CH001782 |
Chromatography Summary: | Phospholipid protocol |
Instrument Name: | Thermo Dionex Ultimate 3000 |
Column Name: | Phenomenex Luna Silica (150 x 2.0mm,3um) |
Chromatography Type: | Normal phase |
Chromatography ID: | CH001783 |
Chromatography Summary: | Triacylglycerols |
Instrument Name: | Thermo Dionex Ultimate 3000 |
Column Name: | Phenomenex Luna Silica (150 x 1.0mm,3um) |
Chromatography Type: | Reversed phase |
MS:
MS ID: | MS002262 |
Analysis ID: | AN002425 |
Instrument Name: | Thermo Fusion Orbitrap |
Instrument Type: | Orbitrap |
MS Type: | ESI |
MS Comments: | Analysis of LC/MS data was performed using software package Compound Discoverer (ThermoFisher Scientific) with an in-house generated analysis workflow and oxidized phospholipid database. |
Ion Mode: | NEGATIVE |
MS ID: | MS002263 |
Analysis ID: | AN002426 |
Instrument Name: | Thermo Q Exactive HF hybrid Orbitrap |
Instrument Type: | Orbitrap |
MS Type: | ESI |
MS Comments: | TAG cations were formed through molecular ammonium adduction (+NH4). Analysis was performed at a resolution of 140,000 for the full MS scan and 17,500 for the MS2 scan in a data-dependent mode. The scan range for MS analysis was 300–1200 m/z with a maximum injection time of 128 ms using one microscan. A maximum injection time of 500 ms was used for MS2 (high energy collisional dissociation (HCD)) analysis with collision energy set to 24. An isolation window of 1.0 Da was set for the MS and MS2 scans. Capillary spray voltage was set at 4.5 kV, and capillary temperature was 320 °C. Sheath gas was set to eight arbitrary units and the S-lens Rf level was set to 60. Standards for TAGs and their oxygenated metabolites were from Avanti Polar Lipids and Cayman Chemicals. |
Ion Mode: | POSITIVE |