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MB Sample ID: SA205263
| Local Sample ID: | C8_6 |
| Subject ID: | SU002222 |
| Subject Type: | Mammal |
| Subject Species: | Mus musculus |
| Taxonomy ID: | 10090 |
Select appropriate tab below to view additional metadata details:
Subject:
| Subject ID: | SU002222 |
| Subject Type: | Mammal |
| Subject Species: | Mus musculus |
| Taxonomy ID: | 10090 |
Factors:
| Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
|---|---|---|---|---|
| C8_6 | SA205263 | FL025173 | Control | pheotype |
Collection:
| Collection ID: | CO002215 |
| Collection Summary: | TKO p53LSL R172H/+ Dicer1flox/flox Ptenflox/flox Amhr2cre/+ mice were generated by mating p53LSL-R172H/+Dicer1flox/floxPtenflox/flox female mice with Dicer1flox/floxPtenflox/floxAmhr2cre/+ male mice. For TKO controls, p53LSL-R172H/+Dicer1flox/floxPtenflox/flox were generated. TKO control mice carry the same genetic makeup as TKO mice but do not develop HGSC. A sequential serum collection protocol was conducted to collect samples from TKO and TKO controls every two weeks starting at 8 weeks of age until a humane end point for sacrifice or development of ascites. TKO mice were sacrificed at Indiana University School of Medicine in accordance with animal protocol (21124) approved by the IACUC. |
| Sample Type: | Blood (serum) |
Treatment:
| Treatment ID: | TR002234 |
| Treatment Summary: | TKO p53LSL R172H/+ Dicer1flox/flox Ptenflox/flox Amhr2cre/+ mice were generated by mating p53LSL-R172H/+Dicer1flox/floxPtenflox/flox female mice with Dicer1flox/floxPtenflox/floxAmhr2cre/+ male mice. For TKO controls, p53LSL-R172H/+Dicer1flox/floxPtenflox/flox were generated. TKO control mice carry the same genetic makeup as TKO mice but do not develop HGSC. A sequential serum collection protocol was conducted to collect samples from TKO and TKO controls every two weeks starting at 8 weeks of age until a humane end point for sacrifice or development of ascites. TKO mice were sacrificed at Indiana University School of Medicine in accordance with animal protocol (21124) approved by the IACUC. |
Sample Preparation:
| Sampleprep ID: | SP002228 |
| Sampleprep Summary: | An extraction solvent consisting of a mixture of isotopically labeled internal standards including 808 µM 13C6 arginine and 212 µM 13C methionine D3 was added to methanol in a 1:60 ratio and stored at 4 °C until further use. Serum samples were thawed on ice, followed by protein precipitation in a 10 µl serum aliquot with extraction solvent in a 1:3 ratio. Samples were vortexed for 30 s and centrifuged at 13,000 rpm for 7 min. To enhance CE peak shape, the resulting supernatant was diluted in a 1:4 ratio with the sample diluent containing 133 mM ammonium acetate, 0.1% formic acid and 1 µM 13C phenylalanine. |
Combined analysis:
| Analysis ID | AN003498 |
|---|---|
| Analysis type | MS |
| Chromatography type | CE |
| Chromatography system | ZipChip (908 Devices) |
| Column | HS chip |
| MS Type | ESI |
| MS instrument type | Orbitrap |
| MS instrument name | Thermo Exactive Plus Orbitrap |
| Ion Mode | POSITIVE |
| Units | Micromolars |
Chromatography:
| Chromatography ID: | CH002584 |
| Instrument Name: | ZipChip (908 Devices) |
| Column Name: | HS chip |
| Chromatography Type: | CE |
MS:
| MS ID: | MS003258 |
| Analysis ID: | AN003498 |
| Instrument Name: | Thermo Exactive Plus Orbitrap |
| Instrument Type: | Orbitrap |
| MS Type: | ESI |
| MS Comments: | µCE-HRMS analyses were performed using the microchip ZipChip® capillary electrophoresis system (classic interface, 908 Devices, Boston MA), coupled to a high-resolution accurate mass Q Exactive plus mass spectrometer (Thermo Fisher Scientific, MA). µCE-HRMS separations used ZipChip® HS chips (10 cm channel). All experiments were performed in positive ionization mode in the 50-500 m/z range at a mass resolution setting of 17,500. The capillary temperature was set to 200 °C and the sheath gas flow rate was 2 psi. The automatic gain control (AGC) target value was set to 3E6 and the maximum injection time was 20 ms. Data were acquired using Xcalibur 3.0 (Thermo Scientific) and were imported to Skyline software14 for peak picking and integration. The peak picking procedure used the analyte accurate m/z and migration time. Peak areas obtained from Skyline were exported as spreadsheets for further analysis. Quantitation was performed with the analyte peak areas relative to the peak area of one of the three isotopically labeled internal standards (13C6 arginine, 13C methionine D3 and 13C phenylalanine) chosen based on migration time similarities |
| Ion Mode: | POSITIVE |
