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MB Sample ID: SA123986
Local Sample ID: | 138 |
Subject ID: | SU001524 |
Subject Type: | Human |
Subject Species: | Homo sapiens |
Taxonomy ID: | 9606 |
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Sample Preparation:
Sampleprep ID: | SP001532 |
Sampleprep Summary: | 4 mL frozen commercial pooled human serum (Innovative Research, Novi, MI, USA) was thawed at 4 °C, vortexed and aliquoted into 50 µL portions in 2 mL Eppendorf vials. Every 50 µL portion was mixed with 250 µL cold methanol and vortexed to precipitate proteins.38 After 20 min incubation at −20 °C, these mixtures were centrifuged at 20,800 x g for 10 min at 4 °C. The supernatants were transferred into clean 2.0 mL Eppendorf vials and then dried in an Eppendorf Vacufuge (Brinkmann Instruments, Westbury, NY, USA). The residue in each Eppendorf vial was reconstituted in 50 µL H2O:ACN (2:3 v/v), vortexed and centrifuged at 20,800 x g for 10 min at 4 °C. The supernatant in all Eppendorf vials was pooled into a 5 mL Eppendorf vial, vortexed and centrifuged at xx,xxx x g (To Dan: I should have used 5000 rpm or a typical rpm in the big centrifuge in the biosample prep lab) for 10 min at 4 °C to further remove any solid residue. The resultant supernatant was aliquoted into 50 µL portions in 1.5 mL Eppendorf vials and stored at −80 °C. Prior to repeated injections to LC-MS in this work, 8 portions were diluted to 200 µL each with H2O:ACN (2:3 v/v), pooled into a 2 mL LC vial and vortexed. |