Metabolomics Workbench : NIH Data Repository

MB Sample ID: SA090249

Local Sample ID:	nHTP_5neg_T21_1-29-5RGW
Subject ID:	SU001309
Subject Type:	Human
Subject Species:	Homo sapiens
Taxonomy ID:	9606
Age Or Age Range:	0.5 - 76.5
Gender:	Male and female

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Sample Preparation:

Sampleprep ID:	SP001317
Sampleprep Summary:	For plasma analyses, a volume of 20μL of was extracted in 480μL of ice-cold methanol:acetonitrile:water (5:3:2). For CSF analyses, a volume of 20μL was extracted in 180μL and for cell-based experiments, 2 million cells were extracted in 1mL of the same ice-cold lysis solution. Subsequently, these solutions were vortexed for 30 minutes at 4°C. Insoluble proteins were pelleted by centrifugation (10 minutes at 4°C and 12,000 g) and supernatants were collected and stored at -80°C until analysis. For quantitative analysis of kynurenine pathway (KP) metabolites, supernatants were spun in a Speedvac until dry and resuspended in 0.1% formic acid in water as previously described (PMID: 30213797, 30143553).

Sampleprep ID:

SP001317

Sampleprep Summary:

For plasma analyses, a volume of 20μL of was extracted in 480μL of ice-cold methanol:acetonitrile:water (5:3:2). For CSF analyses, a volume of 20μL was extracted in 180μL and for cell-based experiments, 2 million cells were extracted in 1mL of the same ice-cold lysis solution. Subsequently, these solutions were vortexed for 30 minutes at 4°C. Insoluble proteins were pelleted by centrifugation (10 minutes at 4°C and 12,000 g) and supernatants were collected and stored at -80°C until analysis. For quantitative analysis of kynurenine pathway (KP) metabolites, supernatants were spun in a Speedvac until dry and resuspended in 0.1% formic acid in water as previously described (PMID: 30213797, 30143553).