Metabolomics Workbench : NIH Data Repository

MB Sample ID: SA174465

Local Sample ID:	146
Subject ID:	SU001941
Subject Type:	Cultured cells
Subject Species:	Homo sapiens
Taxonomy ID:	9606

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Treatment:

Treatment ID:	TR001953
Treatment Summary:	Freshly collected erythrocytes were washed, resuspended to 10% hematocrit in buffer [25mM HEPES (pH 7.4), 120mM NaCl, 5.4mM KCl, 1.8mM CaCl2, and 1mM NaH2PO4], incubated for 1 hour at 37ºC, then centrifuged at 2000xg for 5 minutes at 4ºC, resuspended in fresh wash buffer to 30-40% hematocrit, and split into 210uL aliquots of packed erythrocytes. Erythrocytes were resuspended in 253uL of RPMI containing 11.9mM D-[1,2,3-13C] glucose (Millipore-Sigma) and either treated with POM-SF or POM-HEX at five times the EC50s for MetHb formation, or an untreated solvent control with the balance volume to 550uL using washing buffer. All samples were incubated at 37ºC shaking at 500 RPM. Triplicate samples of supernatants and packed erythrocytes were collected at 0, 0.5, 1, and 6-hour intervals and snap-frozen in liquid N2.

Treatment ID:

TR001953

Treatment Summary:

Freshly collected erythrocytes were washed, resuspended to 10% hematocrit in buffer [25mM HEPES (pH 7.4), 120mM NaCl, 5.4mM KCl, 1.8mM CaCl2, and 1mM NaH2PO4], incubated for 1 hour at 37ºC, then centrifuged at 2000xg for 5 minutes at 4ºC, resuspended in fresh wash buffer to 30-40% hematocrit, and split into 210uL aliquots of packed erythrocytes. Erythrocytes were resuspended in 253uL of RPMI containing 11.9mM D-[1,2,3-13C] glucose (Millipore-Sigma) and either treated with POM-SF or POM-HEX at five times the EC50s for MetHb formation, or an untreated solvent control with the balance volume to 550uL using washing buffer. All samples were incubated at 37ºC shaking at 500 RPM. Triplicate samples of supernatants and packed erythrocytes were collected at 0, 0.5, 1, and 6-hour intervals and snap-frozen in liquid N2.