Metabolomics Workbench : NIH Data Repository

MB Sample ID: SA063953

Local Sample ID:	#B10_0025 YKT_11/8/16_#16
Subject ID:	SU001058
Subject Type:	Mammal
Subject Species:	Mus musculus
Taxonomy ID:	10090
Genotype Strain:	FVB/N
Age Or Age Range:	16.5 to 18.5 weeks old
Weight Or Weight Range:	25 to 30 grams
Gender:	Male
Animal Animal Supplier:	in house breeding
Animal Feed:	Standard chow

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Sample Preparation:

Sampleprep ID:	SP001065
Sampleprep Summary:	Cardiac tissue samples were homogenized and sonicated in phosphate buffered saline (PBS) (pH 7.4). The Bicinchoninic acid (BCA) assay (Thermo Scientific) was used to determine protein concentration. A mixture of internal standards (10µl) in CHCl3: methanol (1:1) and 200µl of CHCl3/methanol (2:1) were added to each sample (ventricles- 50µg protein in 10µl; plasma- 10µl) before being briefly vortexed. Samples were first mixed (rotary mixer, 10 min), sonicated (water bath, 30 min at room temperature [RT]) then rested at RT (20 min). Samples were centrifuged (16,000 g, 10 min) and the supernatant was dried under a constant stream of nitrogen gas at 40°C. Extracted lipids were resuspended in 50µl H2O saturated butanol then sonicated (10 min), before 50 µl of methanol with 10mM ammonium was added. Resuspended extracts were centrifuged (3,350 g, 5 min), and the supernatant transferred to glass vials with Teflon insert caps and stored at -80°C until analyzed. Lipid extracts were thawed at RT for an hour, then sonicated in a water bath for 15 min at RT before analysis.

Sampleprep ID:

SP001065

Sampleprep Summary:

Cardiac tissue samples were homogenized and sonicated in phosphate buffered saline (PBS) (pH 7.4). The Bicinchoninic acid (BCA) assay (Thermo Scientific) was used to determine protein concentration. A mixture of internal standards (10µl) in CHCl3: methanol (1:1) and 200µl of CHCl3/methanol (2:1) were added to each sample (ventricles- 50µg protein in 10µl; plasma- 10µl) before being briefly vortexed. Samples were first mixed (rotary mixer, 10 min), sonicated (water bath, 30 min at room temperature [RT]) then rested at RT (20 min). Samples were centrifuged (16,000 g, 10 min) and the supernatant was dried under a constant stream of nitrogen gas at 40°C. Extracted lipids were resuspended in 50µl H2O saturated butanol then sonicated (10 min), before 50 µl of methanol with 10mM ammonium was added. Resuspended extracts were centrifuged (3,350 g, 5 min), and the supernatant transferred to glass vials with Teflon insert caps and stored at -80°C until analyzed. Lipid extracts were thawed at RT for an hour, then sonicated in a water bath for 15 min at RT before analysis.