Metabolomics Workbench : NIH Data Repository

MB Sample ID: SA343198

Local Sample ID:	C8
Subject ID:	SU003293
Subject Type:	Cultured cells
Subject Species:	Homo sapiens
Taxonomy ID:	9606

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Sample Preparation:

Sampleprep ID:	SP003300
Sampleprep Summary:	Cells without stress treatment or treated with 0.6 M sorbitol stress for 1.5 h were scraped off and centrifuged to collect cell pellets. The cell pellets were thawed on ice, resuspended in stress granule lysis buffer (50 mM Tris, 100 mM K2OAc, 2 mM MgOAc, 0.5 mM DTT, 50 μg/mL heparin, 0.5% NP-40, 0.02% antifoam B, proteinase inhibitor cocktail, pH 7.4) and lysis by syringe. Lysates were centrifuged at 1000 × g for 5 min to collect supernatants. Supernatants were centrifuge at 18000 × g for 20 min to collect pellets and the pellets were washed once with stress granule lysis buffer. Then, pellets were resuspended in stress granule lysis buffer and centrifuged at 850 × g for 2 min to collect supernatants containing stress granule. Then NAD+, ATP and 13C-glucose were added for reaction at 37 'C for 0, 1 and 3 hr. The reaction was quenched by 3-fold methanol and centrifuge to collect the supernant for analysis.

Sampleprep ID:

SP003300

Sampleprep Summary:

Cells without stress treatment or treated with 0.6 M sorbitol stress for 1.5 h were scraped off and centrifuged to collect cell pellets. The cell pellets were thawed on ice, resuspended in stress granule lysis buffer (50 mM Tris, 100 mM K2OAc, 2 mM MgOAc, 0.5 mM DTT, 50 μg/mL heparin, 0.5% NP-40, 0.02% antifoam B, proteinase inhibitor cocktail, pH 7.4) and lysis by syringe. Lysates were centrifuged at 1000 × g for 5 min to collect supernatants. Supernatants were centrifuge at 18000 × g for 20 min to collect pellets and the pellets were washed once with stress granule lysis buffer. Then, pellets were resuspended in stress granule lysis buffer and centrifuged at 850 × g for 2 min to collect supernatants containing stress granule. Then NAD+, ATP and 13C-glucose were added for reaction at 37 'C for 0, 1 and 3 hr. The reaction was quenched by 3-fold methanol and centrifuge to collect the supernant for analysis.