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MB Sample ID: SA013971
| Local Sample ID: | CD3 WT1 042215 |
| Subject ID: | SU000327 |
| Subject Type: | Animal |
| Subject Species: | Mus musculus |
| Taxonomy ID: | 10090 |
| Animal Animal Supplier: | WT (CRBN +/+) mice are from Jackson labs and CRBN -/- mice have been bred at Moffitt Cancer Center |
| Animal Housing: | Standard bedding + cotton pads, hepa filtered air |
| Animal Light Cycle: | Dark 6pm-6am |
| Animal Feed: | Feed from Harlan, not sterile |
| Animal Water: | autoclaved H2O |
| Animal Inclusion Criteria: | Animals used in this experiment were sex-matched and age matched. The ages were 8-12 weeks old in both sets of experiments |
| Species Group: | Mammal |
Select appropriate tab below to view additional metadata details:
Subject:
| Subject ID: | SU000327 |
| Subject Type: | Animal |
| Subject Species: | Mus musculus |
| Taxonomy ID: | 10090 |
| Animal Animal Supplier: | WT (CRBN +/+) mice are from Jackson labs and CRBN -/- mice have been bred at Moffitt Cancer Center |
| Animal Housing: | Standard bedding + cotton pads, hepa filtered air |
| Animal Light Cycle: | Dark 6pm-6am |
| Animal Feed: | Feed from Harlan, not sterile |
| Animal Water: | autoclaved H2O |
| Animal Inclusion Criteria: | Animals used in this experiment were sex-matched and age matched. The ages were 8-12 weeks old in both sets of experiments |
| Species Group: | Mammal |
Factors:
| Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
|---|---|---|---|---|
| CD3 WT1 042215 | SA013971 | FL003231 | CRBN +/+ | Strain |
| CD3 WT1 042215 | SA013971 | FL003231 | anti-CD3 stimulation | Treatment |
Collection:
| Collection ID: | CO000321 |
| Collection Summary: | - |
| Sample Type: | cell |
| Collection Method: | centrifuged in ficoll-gradient to remove debris, then washed in PBS |
| Collection Time: | 0 or 24 hours after stimulation |
| Storage Conditions: | -80°C |
| Collection Vials: | 15mL conical tubes |
| Storage Vials: | microfuge tubes |
Treatment:
| Treatment ID: | TR000341 |
| Treatment Summary: | - |
| Animal Anesthesia: | CO2 asphyxiation |
| Animal Acclimation Duration: | Mice from Jackson are subjected to a week-long acclimation period in our facility prior to experimental use. |
| Animal Endp Tissue Coll List: | Spleen into cell culture media |
| Cell Growth Container: | 96-well round-bottom plate that was coated with anti-CD3 antibody |
| Cell Growth Rate: | Murine T cells generally divide after 48-72 hours of stimulation. |
| Cell Media: | RPMI 1640 with 10% FBS, pen/strep, gentamycin, NEAA, Sodium pyruvate, fungizone (Amphotericin B), 2-mercaptoethanol |
| Cell Envir Cond: | 37°C, normoxic conditions |
Sample Preparation:
| Sampleprep ID: | SP000335 |
| Sampleprep Summary: | - |
| Sampleprep Protocol Filename: | Consolidated Cellular Metabolomics with DNA quant.pdf |
| Sample Spiking: | Appendix A - Internal Standard Prep GLCMS.pdf |
Combined analysis:
| Analysis ID | AN000486 | AN000487 |
|---|---|---|
| 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 area | Peak area |
Chromatography:
| Chromatography ID: | CH000347 |
| Methods Filename: | Metabolomics_LCMSProtocol.pdf |
| Instrument Name: | Thermo Scientific-Dionex Ultimate 3000 |
| Column Name: | ACE Excel 2 C18-PFP (100 x 2.1mm, 2um) |
| Flow Rate: | 0.350mL/min-0.600mL/min |
| Solvent A: | 100% water; 0.1% formic acid |
| Solvent B: | 100% acetonitrile |
| Chromatography Type: | Reversed phase |
MS:
| MS ID: | MS000424 |
| Analysis ID: | AN000486 |
| Instrument Name: | Thermo Q Exactive Orbitrap |
| Instrument Type: | Orbitrap |
| MS Type: | ESI |
| Ion Mode: | POSITIVE |
| Analysis Protocol File: | Metabolomics_LCMSProtocol.pdf |
| MS ID: | MS000425 |
| Analysis ID: | AN000487 |
| Instrument Name: | Thermo Q Exactive Orbitrap |
| Instrument Type: | Orbitrap |
| MS Type: | ESI |
| Ion Mode: | NEGATIVE |
| Analysis Protocol File: | Metabolomics_LCMSProtocol.pdf |
