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MB Sample ID: SA095498
Local Sample ID: | HeLa-wnt-20min-02 |
Subject ID: | SU001394 |
Subject Type: | Cultured cells |
Subject Species: | Homo sapiens |
Taxonomy ID: | 9606 |
Select appropriate tab below to view additional metadata details:
Subject:
Subject ID: | SU001394 |
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 |
---|---|---|---|---|
HeLa-wnt-20min-02 | SA095498 | FL013554 | wnt | Treatment |
HeLa-wnt-20min-02 | SA095498 | FL013554 | 20 | Treatment_Time(min) |
Collection:
Collection ID: | CO001389 |
Collection Summary: | Extraction began by placing the 6-well dishes on ice and washing each well with 2mL of a 150 mM ammonium acetate solution (pH 7.4, 4°C). Post wash, metabolites were extracted from each well on dry ice with 500 μL of a (1:4) dH20, MeOH solution at -80°C. The samples were then incubated for 15 min at -80°C. Afterwards, the cells were scraped into solutions which were then transferred to eppitubes and spun down at 4°C for 10 min at 17,000g. The supernatants were transferred to new glass vials and dried with an EZ2-Elite lyophilizer (Genevac). These dried metabolites were then stored at -80°C until they could be run on the LC/MS. |
Sample Type: | Cultured cells |
Storage Conditions: | Described in summary |
Treatment:
Treatment ID: | TR001409 |
Treatment Summary: | The cells were incubated with DMEM(no glucose) supplemented with 10% dialyzed FBS, 1% P/S, and 10 mM UC13-glucose with either vehicle (0.1% BSA in dH20) or wnt (100ng/mL) added. Cells were treated for either 20 min or 60 min with each treatment group having a vehicle matched time control. |
Sample Preparation:
Sampleprep ID: | SP001402 |
Sampleprep Summary: | Extraction began by placing the 6-well dishes on ice and washing each well with 2mL of a 150 mM ammonium acetate solution (pH 7.4, 4°C). Post wash, metabolites were extracted from each well on dry ice with 500 μL of a (1:4) dH20, MeOH solution at -80°C. The samples were then incubated for 15 min at -80°C. Afterwards, the metabolite solutions were transferred to eppitubes and spun down at 4°C for 10 min at 17,000g. The supernatants were transferred to new vials and dried with an EZ2-Elite lyophilizer (Genevac). These dried metabolites were then stored at -80°C until they could be run on the LC/MS. |
Extract Storage: | -80℃ |
Combined analysis:
Analysis ID | AN002196 |
---|---|
Analysis type | MS |
Chromatography type | HILIC |
Chromatography system | Thermo Vanquish |
Column | Phenomenex Luna NH2 (150 x 2.1mm,3um) |
MS Type | ESI |
MS instrument type | Orbitrap |
MS instrument name | Thermo Q Exactive Orbitrap |
Ion Mode | UNSPECIFIED |
Units | MS1 AUC integration |
Chromatography:
Chromatography ID: | CH001610 |
Chromatography Summary: | Samples were run on a Vanquish (Thermo Scientific) UHPLC system with mobile phase A (5mM NH4AcO, pH 9.9) and mobile phase B (ACN) at a flow rate of 200 µL/min on a Luna 3 um NH2 100A (150 × 2.0mm) at 40°C with a gradient going from 15% A to 95% A in 18 min followed by an 11 minute isocratic step. |
Instrument Name: | Thermo Vanquish |
Column Name: | Phenomenex Luna NH2 (150 x 2.1mm,3um) |
Column Temperature: | 40 |
Flow Gradient: | 15% A to 95% A in 18 min followed by an 11 minute isocratic step |
Flow Rate: | 200 µL/min |
Solvent A: | 100% water; 5 mM ammonium acetate, pH 9.9 |
Solvent B: | 100% acetonitrile |
Chromatography Type: | HILIC |
MS:
MS ID: | MS002042 |
Analysis ID: | AN002196 |
Instrument Name: | Thermo Q Exactive Orbitrap |
Instrument Type: | Orbitrap |
MS Type: | ESI |
MS Comments: | Method runs in polarity switching mode. Features were assigned via MzMine 2 and an in house list of MS1 and RT times. |
Ion Mode: | UNSPECIFIED |