Metabolomics Workbench : NIH Data Repository

MB Sample ID: SA212091

Local Sample ID:	MS56-011
Subject ID:	SU002309
Subject Type:	Mammal
Subject Species:	Mus musculus
Taxonomy ID:	10090

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Sample Preparation:

Sampleprep ID:	SP002315
Sampleprep Summary:	For tissue extraction, samples were homogenized in metabolite extraction buffer using the proportion 25 μl/mg of buffer with Precellys Lysing tubes (Bertin Instruments). After that, extracts were kept in the freezer overnight and the following day centrifuged twice at max speed at 4C˚ to remove the protein precipitates. Equal volumes of supernatants were spiked in with 13C arginine (Cambridge Isotopes) for quantification of arginine content. For extraction of the tissue interstitial for interstitial fluid extraction, the organ was washed in saline solution and then a portion was centrifuged at for 10 min at 4°C at 106 x g using 20 µm nylon filters (Spectrum Labs, Waltham, MA, 148134) affixed on top of 2 ml Eppendorf tubes. 1μl of the eluate was extracted in 45μl of extraction buffer and frozen overnight. The following day, all extracted were centrifuged twice at max speed at 4C˚to remove the protein precipitates. Supernatants were finally spiked in with 13C arginine (Cambridge Isotopes) for arginine quantification.

Sampleprep ID:

SP002315

Sampleprep Summary:

For tissue extraction, samples were homogenized in metabolite extraction buffer using the proportion 25 μl/mg of buffer with Precellys Lysing tubes (Bertin Instruments). After that, extracts were kept in the freezer overnight and the following day centrifuged twice at max speed at 4C˚ to remove the protein precipitates. Equal volumes of supernatants were spiked in with 13C arginine (Cambridge Isotopes) for quantification of arginine content. For extraction of the tissue interstitial for interstitial fluid extraction, the organ was washed in saline solution and then a portion was centrifuged at for 10 min at 4°C at 106 x g using 20 µm nylon filters (Spectrum Labs, Waltham, MA, 148134) affixed on top of 2 ml Eppendorf tubes. 1μl of the eluate was extracted in 45μl of extraction buffer and frozen overnight. The following day, all extracted were centrifuged twice at max speed at 4C˚to remove the protein precipitates. Supernatants were finally spiked in with 13C arginine (Cambridge Isotopes) for arginine quantification.