Return to study ST001345 main page
MB Sample ID: SA097843
Local Sample ID: | TLR4IFNG_IFNARKO_2 |
Subject ID: | SU001419 |
Subject Type: | Cultured cells |
Subject Species: | Mus musculus |
Taxonomy ID: | 10090 |
Genotype Strain: | C57BL6/J |
Age Or Age Range: | 8-12 |
Gender: | Male |
Cell Passage Number: | Primary |
Select appropriate tab below to view additional metadata details:
Subject:
Subject ID: | SU001419 |
Subject Type: | Cultured cells |
Subject Species: | Mus musculus |
Taxonomy ID: | 10090 |
Genotype Strain: | C57BL6/J |
Age Or Age Range: | 8-12 |
Gender: | Male |
Cell Passage Number: | Primary |
Factors:
Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
---|---|---|---|---|
TLR4IFNG_IFNARKO_2 | SA097843 | FL013721 | TLR4IFNG | Treatment |
TLR4IFNG_IFNARKO_2 | SA097843 | FL013721 | IFNAR-/- | Genotype |
Collection:
Collection ID: | CO001414 |
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: | TR001434 |
Treatment Summary: | WT or IFNAR deficient BMDMs were seeded at a density of 9x105 cell/well in 6 well dishes (Fisher 08-772-1B). On day 8, BMDM were stimulated with IFNB, TLR3, IFNG, TLR4, and TLR4+IFNg ligands in 5% FBS BMDM media for 48 h. |
Sample Preparation:
Sampleprep ID: | SP001427 |
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 | AN002239 |
---|---|
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: | CH001645 |
Instrument Name: | none |
Column Name: | none |
Chromatography Type: | None (Direct infusion) |
MS:
MS ID: | MS002085 |
Analysis ID: | AN002239 |
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 |