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MB Sample ID: SA114117
Local Sample ID: | LIPPOS17MAR_47.1 |
Subject ID: | SU001479 |
Subject Type: | Cultured cells |
Subject Species: | Homo sapiens |
Taxonomy ID: | 9606 |
Select appropriate tab below to view additional metadata details:
Subject:
Subject ID: | SU001479 |
Subject Type: | Cultured cells |
Subject Species: | Homo sapiens |
Taxonomy ID: | 9606 |
Factors:
Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
---|---|---|---|---|
LIPPOS17MAR_47.1 | SA114117 | FL014381 | Untreated | Treatment_Status |
LIPPOS17MAR_47.1 | SA114117 | FL014381 | Higher | MDM2 Amplification |
LIPPOS17MAR_47.1 | SA114117 | FL014381 | 141 | Cell line |
Collection:
Collection ID: | CO001474 |
Collection Summary: | Cells were collected using 1mL of cold (-20C) methanol and a cell scraper and then immediately stored at -80C. |
Sample Type: | Cultured cells |
Treatment:
Treatment ID: | TR001494 |
Treatment Summary: | DDLPS were plated in 60mm dishes and drug treated with atorvastatin for 72 hours.All cells were cultured in Dulbecco's modified Eagle's medium (DMEM) and supplemented with 10\% fetal bovine serum (FBS), 100 U/mL penicillin, and 100 U/mL streptomycin. These cells were cultured in a humidified chamber delivering 5% CO2 at 37 degrees C. |
Treatment Compound: | Atorvastatin |
Cell Media: | Dulbecco's modified Eagle's medium |
Sample Preparation:
Sampleprep ID: | SP001487 |
Sampleprep Summary: | Cells were pelleted and processed using a chloroform:methanol homogenization followed by an isoproponal:acetonitrile extraction. Samples were separated by reverse phase HPLC using a Prominence 20 UFLCXR system (Shimadzu, Columbia MD) with a Waters (Milford, MA) CSH C18 column (100mm x 2.1mm 1.7 um particle size) maintained at 55C and a 20 minute aqueous/acetonitrile/isopropanol gradient, at a flow rate of 225 ul/min. For electrospray ionization positive mode, Solvent A was 40% water, 60% acetonitrile with 10mM ammonium formate and 0.1% formic acid, and Solvent B was 90% isopropanol, 10% acetonitrile with 10mM ammonium formate and 0.1% formic acid. For electrospray ionization negative mode, Solvent A was 40% water, 60% isopropanol with 10 mM ammonium acetate, and solvent B was 90% isopropanol, 10% acetonitrile with 10 mM ammonium acetate. The initial conditions were 60% A and 40% B, increasing to 43% B at 2 minutes, 50% B at 2.1 minutes., 54% B at 12 minutes, 70% B at 12.1 minutes and 99% B at 18 minutes, held at 99% B until 20.0 minutes before returning to the initial conditions. |
Processing Storage Conditions: | -80℃ |
Extract Storage: | -80℃ |
Combined analysis:
Analysis ID | AN002347 | AN002348 |
---|---|---|
Analysis type | MS | MS |
Chromatography type | Reversed phase | Reversed phase |
Chromatography system | Shimadzu Prominence 20 UFLCXR | Shimadzu Prominence 20 UFLCXR |
Column | Waters XBridge C18 (50 x 2.1mm,3um) | Waters XBridge C18 (50 x 2.1mm,3um) |
MS Type | ESI | ESI |
MS instrument type | Triple TOF | Triple TOF |
MS instrument name | ABI Sciex 5600 TripleTOF | ABI Sciex 5600 TripleTOF |
Ion Mode | POSITIVE | NEGATIVE |
Units | ppm | ppm |
Chromatography:
Chromatography ID: | CH001720 |
Chromatography Summary: | Samples were separated by reverse phase HPLC using a Prominence 20 UFLCXR system (Shimadzu, Columbia MD) with a Waters (Milford, MA) CSH C18 column (100mm x 2.1mm 1.7 um particle size) maintained at 55C and a 20 minute aqueous/acetonitrile/isopropanol gradient, at a flow rate of 225 ul/min. For electrospray ionization positive mode, Solvent A was 40% water, 60% acetonitrile with 10mM ammonium formate and 0.1% formic acid, and Solvent B was 90% isopropanol, 10% acetonitrile with 10mM ammonium formate and 0.1% formic acid. For electrospray ionization negative mode, Solvent A was 40% water, 60% isopropanol with 10 mM ammonium acetate, and solvent B was 90% isopropanol, 10% acetonitrile with 10 mM ammonium acetate. The initial conditions were 60% A and 40% B, increasing to 43% B at 2 minutes, 50% B at 2.1 minutes, 54% B at 12 minutes, 70% B at 12.1 minutes and 99% B at 18 minutes, held at 99% B until 20.0 minutes before returning to the initial conditions. |
Instrument Name: | Shimadzu Prominence 20 UFLCXR |
Column Name: | Waters XBridge C18 (50 x 2.1mm,3um) |
Column Temperature: | 55 |
Flow Gradient: | The initial conditions were 60% A and 40% B, increasing to 43% B at 2 minutes, 50% B at 2.1 minutes, 54% B at 12 minutes, 70% B at 12.1 minutes and 99% B at 18 minutes, held at 99% B until 20.0 minutes before returning to the initial conditions. |
Flow Rate: | 225 ul/min |
Solvent A: | Pos mode: 40% water/60% acetonitrile; 0.1% formic acid; 10mM ammonium formate, Neg mode: 40% water/60% isopropanol; 10 mM ammonium acetate |
Solvent B: | Pos mode: 90% isopropanol/10% acetonitrile; 0.1% formic acid; 10mM ammonium formate, Neg mode: 90% isopropanol/10% acetonitrile; 10 mM ammonium acetate |
Chromatography Type: | Reversed phase |
MS:
MS ID: | MS002189 |
Analysis ID: | AN002347 |
Instrument Name: | ABI Sciex 5600 TripleTOF |
Instrument Type: | Triple TOF |
MS Type: | ESI |
MS Comments: | The capillary voltage was set at 5.5 kV, with a declustering potential of 80V. The mass spectrometer was operated in IDA (Information Dependent Acquisition) mode with a 100 ms survey scan from 100 to 1200 m/z, and up to 20 MS/MS product ion scans (100 ms) per duty cycle using a collision energy of 50V with a 20V spread. |
Ion Mode: | POSITIVE |
MS ID: | MS002190 |
Analysis ID: | AN002348 |
Instrument Name: | ABI Sciex 5600 TripleTOF |
Instrument Type: | Triple TOF |
MS Type: | ESI |
MS Comments: | The capillary voltage was set at 4.5 kV, with a declustering potential of 80V. The mass spectrometer was operated in IDA (Information Dependent Acquisition) mode with a 100 ms survey scan from 100 to 1200 m/z, and up to 20 MS/MS product ion scans (100 ms) per duty cycle using a collision energy of 50V with a 20V spread. |
Ion Mode: | NEGATIVE |