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MB Sample ID: SA097657
| Local Sample ID: | TLR3+ACLDL_1 |
| Subject ID: | SU001415 |
| Subject Type: | Cultured cells |
| Subject Species: | Mus musculus |
| Taxonomy ID: | 10090 |
| Genotype Strain: | C57BL6/J |
| Age Or Age Range: | 8-12 |
| Gender: | Male |
Select appropriate tab below to view additional metadata details:
Subject:
| Subject ID: | SU001415 |
| Subject Type: | Cultured cells |
| Subject Species: | Mus musculus |
| Taxonomy ID: | 10090 |
| Genotype Strain: | C57BL6/J |
| Age Or Age Range: | 8-12 |
| Gender: | Male |
Factors:
| Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
|---|---|---|---|---|
| TLR3+ACLDL_1 | SA097657 | FL013674 | TLR3+ACLDL | Treatment |
Collection:
| Collection ID: | CO001410 |
| Collection Summary: | Day 6 BMDMs were seeded at a density of 9x105 cell/well in 6 well dishes (Fisher 08-772-1B). On day 8, BMDM were mock-treated (media change) or co-treated with ligands as described below in 5% BMDM media for 48 h. Prior to collection, cell number was assessed by imaging as described below. After removal of culturing media, 1 mL of ice-cold PBS was added into each well and cells were scraped with cell lifters and spun down in glass tubes at 365g for 5 min at 4°C. |
| Sample Type: | Macrophages |
Treatment:
| Treatment ID: | TR001430 |
| Treatment Summary: | BMDMs were seeded at a density of 9x105 cell/well in 6 well dishes (Fisher 08-772-1B). On day 8, BMDM were first pre-loaded with 50 ug/mL of acetylated-LDL for 2 h, followed by TLR2, TLR3, TLR4, TLR7, or TLR9 ligands stimulation in 5% FBS BMDM media for 48 h. |
Sample Preparation:
| Sampleprep ID: | SP001423 |
| Sampleprep Summary: | A modified Bligh and Dyer extraction (Bligh and Dyer, 1959) was carried out on samples. Prior to biphasic extraction, a 13-lipid class Lipidyzer Internal Standard Mix is added to each sample (AB Sciex, 5040156). Following two successive extractions, pooled organic layers were dried down in a Genevac EZ-2 Elite. Lipid samples were resuspended in 1:1 methanol/dichloromethane with 10 mM Ammonium Acetate and transferred to robovials (Thermo 10800107) for analysis. |
Combined analysis:
| Analysis ID | AN002235 |
|---|---|
| Analysis type | MS |
| Chromatography type | None (Direct infusion) |
| Chromatography system | none |
| Column | none |
| MS Type | ESI |
| MS instrument type | Triple quadrupole |
| MS instrument name | ABI Sciex 5500 QTRAP with SelexION |
| Ion Mode | UNSPECIFIED |
| Units | nmol per 10e7 cells |
Chromatography:
| Chromatography ID: | CH001641 |
| Instrument Name: | none |
| Column Name: | none |
| Chromatography Type: | None (Direct infusion) |
MS:
| MS ID: | MS002081 |
| Analysis ID: | AN002235 |
| Instrument Name: | ABI Sciex 5500 QTRAP with SelexION |
| Instrument Type: | Triple quadrupole |
| MS Type: | ESI |
| MS Comments: | Lipidyzer Workflow Manager 1.0 Samples are normalized to cell count |
| Ion Mode: | UNSPECIFIED |
