Metabolomics Workbench : NIH Data Repository

MB Sample ID: SA330420

Local Sample ID:	Serum3DWP4positive
Subject ID:	SU003158
Subject Type:	Mammal
Subject Species:	Mus musculus
Taxonomy ID:	10090
Gender:	Female

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Sample Preparation:

Sampleprep ID:	SP003164
Sampleprep Summary:	To extract metabolites, frozen kidney cortices were weighed, and ca 20 mg of each cortex was added to a tube containing 650 μL of 80% methanol (diluted in dH2O). Sample tubes were placed in a bead homogenizer for 1 minute at a frequency of 30/s. Samples were then placed in a -20℃ freezer for 10 minutes, centrifuged for 10 minutes at 15,000 RPM in 4℃, and the supernatant was moved to a new tube. The remaining pellets were then re-extracted with 400 μL of 80% methanol, following the above procedure of homogenization and centrifugation. The supernatant was then added to the supernatant pool obtained from the first extraction. The pellet was re-extracted for a third time using 350 μL of 80% MeOH and supernatant was again added to the pool from the first two extractions. The supernatant pool was placed at -20℃ for 15 minutes, centrifuged for 20 minutes at 15,000 RPM, and transferred to a new tube. The methanolic extract samples were speed-vacuumed and reconstituted in 70% acetonitrile (diluted in dH2O) containing 0.025% acetic acid at a protein concentration of 3μg/μL. For serum preparation, samples were diluted 10-fold with 70% acetonitrile (diluted in dH2O) containing 0.025% acetic acid, and centrifuged for 20 minutes at 20,000 g.

Sampleprep ID:

SP003164

Sampleprep Summary:

To extract metabolites, frozen kidney cortices were weighed, and ca 20 mg of each cortex was added to a tube containing 650 μL of 80% methanol (diluted in dH2O). Sample tubes were placed in a bead homogenizer for 1 minute at a frequency of 30/s. Samples were then placed in a -20℃ freezer for 10 minutes, centrifuged for 10 minutes at 15,000 RPM in 4℃, and the supernatant was moved to a new tube. The remaining pellets were then re-extracted with 400 μL of 80% methanol, following the above procedure of homogenization and centrifugation. The supernatant was then added to the supernatant pool obtained from the first extraction. The pellet was re-extracted for a third time using 350 μL of 80% MeOH and supernatant was again added to the pool from the first two extractions. The supernatant pool was placed at -20℃ for 15 minutes, centrifuged for 20 minutes at 15,000 RPM, and transferred to a new tube. The methanolic extract samples were speed-vacuumed and reconstituted in 70% acetonitrile (diluted in dH2O) containing 0.025% acetic acid at a protein concentration of 3μg/μL. For serum preparation, samples were diluted 10-fold with 70% acetonitrile (diluted in dH2O) containing 0.025% acetic acid, and centrifuged for 20 minutes at 20,000 g.