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MB Sample ID: SA325751
Local Sample ID: | G2.3 |
Subject ID: | SU003102 |
Subject Type: | Mammal |
Subject Species: | Mus musculus |
Taxonomy ID: | 10090 |
Select appropriate tab below to view additional metadata details:
Subject:
Subject ID: | SU003102 |
Subject Type: | Mammal |
Subject Species: | Mus musculus |
Taxonomy ID: | 10090 |
Factors:
Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
---|---|---|---|---|
G2.3 | SA325751 | FL038739 | Npm1-knockout | Genotype |
Collection:
Collection ID: | CO003095 |
Collection Summary: | BMDMs (1×10^7) were induced toward the reparative phenotype using recombinant murine IL-4.BMDMs were counted and washed with PBS. |
Sample Type: | Bone marrow |
Treatment:
Treatment ID: | TR003111 |
Treatment Summary: | mouse bone marrow cells were flushed out from mouse femurs using RPMI 1640 and cultured in a complete RPMI 1640 medium (added with 10% fetal bovine serum and 1% penicillin/streptomycin) supplemented with 20 ng/ml M-CSF for 7 days to generate mature BMDMs. BMDMs were stimulated with 20 ng/ml IL-4 for 12 h to induce a reparative phenotype. |
Sample Preparation:
Sampleprep ID: | SP003108 |
Sampleprep Summary: | For each sample, 500 μl precooled MeOH/H2O (3/1, v/v) was added in. The samples were then vortexed for 30 sec. After precooling in dry ice, the samples were frozen and thawed three times in liquid nitrogen. They were then vortexed for 30 sec and sonicated for 15 min in an ice-water bath. Next, as a subsequent step, the samples were incubated at -40 °C for 1 h and centrifuged at 12000 rpm for 15 min at 4 °C (RCF = 13800 g, R = 8.6 cm). Afterwards, an aliquot of 400 μl of clear supernatant was collected and dried by spinning. As a reconstitution solution, mix 200 ml of ultrapure water with the residue. The reconstituted samples were vortexed before passing through the filter of the centrifuge tube, after which they were transferred to inserts in injection vials for HPLC-MS/MS analysis. |
Combined analysis:
Analysis ID | AN004909 |
---|---|
Analysis type | MS |
Chromatography type | Ion exchange |
Chromatography system | Thermo Scientific Dionex ICS-6000 HPIC |
Column | Dionex IonPac AS11-HC (250 × 2mm, 9um, 2000Ã) and AG11-HC (50 × 2 mm, 13um) |
MS Type | ESI |
MS instrument type | Triple quadrupole |
MS instrument name | ABI Sciex 6500+ QTrap |
Ion Mode | UNSPECIFIED |
Units | pmol |
Chromatography:
Chromatography ID: | CH003704 |
Instrument Name: | Thermo Scientific Dionex ICS-6000 HPIC |
Column Name: | Dionex IonPac AS11-HC (250 × 2mm, 9um, 2000Ã) and AG11-HC (50 × 2 mm, 13um) |
Column Temperature: | 30 ℃ |
Flow Gradient: | 0.15 mL/min: 0min:20%/80%, 3.01min:20%/80%,12min:50%/50%, 21min:100%/100%, 26min:100%/100%, 26.1min:20%/80%,0min:80%/20%, 30min:80%/20% |
Flow Rate: | 0.15 mL/min |
Solvent A: | 100 mM NaOH/water; water |
Solvent B: | water |
Chromatography Type: | Ion exchange |
Solvent C: | 2 mM acetic acid in methanol |
MS:
MS ID: | MS004652 |
Analysis ID: | AN004909 |
Instrument Name: | ABI Sciex 6500+ QTrap |
Instrument Type: | Triple quadrupole |
MS Type: | ESI |
MS Comments: | AB SCIEX Analyst Work Station Software (1.6.3 AB SCIEX), MultiQuant 3.0.3.software and Chromeleon7 were employed for MRM data acquisition and processing. |
Ion Mode: | UNSPECIFIED |