Return to study ST001320 main page

MB Sample ID: SA095494

Local Sample ID:HeLa-veh-60min-01
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 IDMB Sample IDFactor Level IDLevel ValueFactor Name
HeLa-veh-60min-01SA095494FL013553vehicleTreatment
HeLa-veh-60min-01SA095494FL01355360Treatment_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
  logo