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MB Sample ID: SA314466
| Local Sample ID: | LAT1-4F2hc_WT_delipidated_1 |
| Subject ID: | SU002988 |
| Subject Type: | Cultured cells |
| Subject Species: | Homo sapiens |
| Taxonomy ID: | 9606 |
| Cell Strain Details: | HEK293F |
Select appropriate tab below to view additional metadata details:
Subject:
| Subject ID: | SU002988 |
| Subject Type: | Cultured cells |
| Subject Species: | Homo sapiens |
| Taxonomy ID: | 9606 |
| Cell Strain Details: | HEK293F |
Factors:
| Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
|---|---|---|---|---|
| LAT1-4F2hc_WT_delipidated_1 | SA314466 | FL037442 | Delipidated | Treatment |
Collection:
| Collection ID: | CO002981 |
| Collection Summary: | The LAT1-4F2hc complex was buffer-exchanged into 1M ammonium acetate, pH 7.0 with 2 mM OGNG and incubated with 1 µg trypsin overnight at 37 °C. The digested peptide/lipid mixture was dried using a SpeedVac vacuum concentrator (Thermo Fisher Scientific) |
| Sample Type: | HEK cells |
Treatment:
| Treatment ID: | TR002997 |
| Treatment Summary: | The LAT1-4F2hc complex was buffer-exchanged into 1M ammonium acetate (pH 7.0) with 20 mM OGNG using a 100 kD MWCO centrifugal filter (Amicon Ultra-0.5 ml, Millipore) and incubated for 4 h at room temperature with gentle mixing. Excess lipomicelles and detergents were removed by buffer-exchanging into 1 M ammonium acetate with 2 mM OGNG using the same filter. |
Sample Preparation:
| Sampleprep ID: | SP002994 |
| Sampleprep Summary: | The lipid mixture was reconstituted with 70% mobile phase A (acetonitrile/H2O: 60/40, 10 mM ammonium formate and 0.1% formic acid) and 30% mobile phase B (isopropanol/acetonitrile: 90/10, 10 mM ammonium formate and 0.1% formic acid) for the following LC-MS/MS analysis. |
Combined analysis:
| Analysis ID | AN004712 |
|---|---|
| Analysis type | MS |
| Chromatography type | Reversed phase |
| Chromatography system | Thermo Dionex Ultimate 3000 |
| Column | Thermo Acclaim PepMap 100 C18 (150 x 0.007 mm, 3um, 100A) |
| MS Type | ESI |
| MS instrument type | Orbitrap |
| MS instrument name | Thermo LTQ XL |
| Ion Mode | NEGATIVE |
| Units | Peak area |
Chromatography:
| Chromatography ID: | CH003548 |
| Chromatography Summary: | Column information: https://www.thermofisher.com/order/catalog/product/164568 |
| Instrument Name: | Thermo Dionex Ultimate 3000 |
| Column Name: | Thermo Acclaim PepMap 100 C18 (150 x 0.007 mm, 3um, 100A) |
| Column Temperature: | 40 |
| Flow Gradient: | 30%-99% |
| Flow Rate: | 300 nl/min |
| Solvent A: | 60% acetonitrile/40% water; 10 mM ammonium formate; 0.1% formic acid |
| Solvent B: | 90% isopropanol/10% acetonitrile; 10 mM ammonium formate; 0.1% formic acid |
| Chromatography Type: | Reversed phase |
MS:
| MS ID: | MS004458 |
| Analysis ID: | AN004712 |
| Instrument Name: | Thermo LTQ XL |
| Instrument Type: | Orbitrap |
| MS Type: | ESI |
| MS Comments: | - |
| Ion Mode: | NEGATIVE |
