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MB Sample ID: SA013088
| Local Sample ID: | 18_2 |
| Subject ID: | SU000306 |
| Subject Type: | Animal cells |
| Subject Species: | Mus musculus |
| Taxonomy ID: | 10090 |
| Genotype Strain: | C57/Bl6 |
| Subject Comments: | Primary mouse myocytes were used for this study. The cells were obtained from gastrocnemius muscle of male 8 weeks-old C57BL/6J mice. |
| Species Group: | Mammal |
Select appropriate tab below to view additional metadata details:
Subject:
| Subject ID: | SU000306 |
| Subject Type: | Animal cells |
| Subject Species: | Mus musculus |
| Taxonomy ID: | 10090 |
| Genotype Strain: | C57/Bl6 |
| Subject Comments: | Primary mouse myocytes were used for this study. The cells were obtained from gastrocnemius muscle of male 8 weeks-old C57BL/6J mice. |
| Species Group: | Mammal |
Factors:
| Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
|---|---|---|---|---|
| 18_2 | SA013088 | FL003100 | CLFS | Treatment |
Collection:
| Collection ID: | CO000300 |
| Collection Summary: | Conditioned media was collected from the cell culture dish, and centrifuged at 3000xg, 4¡C for 10 min to remove cell debris. The supernatant was transferred into new tube and stored at -80¡C. For the metabolomics analysis, 500 uL of samples was put into 1.5 mL tube. |
| Sample Type: | Cells |
Treatment:
| Treatment ID: | TR000320 |
| Treatment Summary: | Primary mouse myocytes were differentiated into myotubes for 3 days, and electrically stimulated at 2.5V, 1.0 Hz by C-PAGE EP (Ionoptix). Conditioned media was collected every 24 hr of stimulation. Conditioned media from the cells without electrical stimulation was also collected as a negative control. |
Sample Preparation:
| Sampleprep ID: | SP000314 |
| Sampleprep Summary: | see Sample Preparation of Cell Lysates.pdf |
| Sampleprep Protocol Filename: | Sample_Preparation_of_Cell_Lysates.pdf |
Combined analysis:
| Analysis ID | AN000454 | AN000455 |
|---|---|---|
| Analysis type | MS | MS |
| Chromatography type | Reversed phase | Reversed phase |
| Chromatography system | Thermo Scientific-Dionex Ultimate 3000: | Thermo Scientific-Dionex Ultimate 3000: |
| Column | ACE Excel 2 C18-PFP (100 x 2.1mm, 2um) | ACE Excel 2 C18-PFP (100 x 2.1mm, 2um) |
| MS Type | ESI | ESI |
| MS instrument type | Orbitrap | Orbitrap |
| MS instrument name | Thermo Q Exactive Orbitrap | Thermo Q Exactive Orbitrap |
| Ion Mode | POSITIVE | NEGATIVE |
| Units | peak height | peak height |
Chromatography:
| Chromatography ID: | CH000324 |
| Chromatography Comments: | UHPLC, Thermo Scientific-Dionex Ultimate 3000: While setting up sequence, ensure that method to be utilized is PFP-metabolomics-neg350-NoBuffer-0-SID and PFP-metabolomics-neg-350-dd-MS2 for negative sequences. For positive sequences utilize PFP-metabolomics-pos-350-0-SID and PFP-metabolomics-pos-350-dd-MS2. Check the lines for air bubbles and purge line if present. Set injection volume to 2uL for positive samples and 4uL for negative samples. |
| Instrument Name: | Thermo Scientific-Dionex Ultimate 3000: |
| Column Name: | ACE Excel 2 C18-PFP (100 x 2.1mm, 2um) |
| Chromatography Type: | Reversed phase |
MS:
| MS ID: | MS000394 |
| Analysis ID: | AN000454 |
| Instrument Name: | Thermo Q Exactive Orbitrap |
| Instrument Type: | Orbitrap |
| MS Type: | ESI |
| Ion Mode: | POSITIVE |
| Analysis Protocol File: | ST286_Metabolomics_LCMSProtocol.pdf |
| MS ID: | MS000395 |
| Analysis ID: | AN000455 |
| Instrument Name: | Thermo Q Exactive Orbitrap |
| Instrument Type: | Orbitrap |
| MS Type: | ESI |
| Ion Mode: | NEGATIVE |
| Analysis Protocol File: | ST286_Metabolomics_LCMSProtocol.pdf |
