The levels of 2DG6P in treated and untreated EAE with 2DG were measured in the spinal cord by ultrahigh performance liquid chromatography-tandem mass spectroscopy (UPLC-MS/MS) (Waters) as described in SI Appendix . The glucose and lactate levels in media were measured using Glucose Glo and Lactate Glo assay kits (Promega) as per the manufacturer's instructions. In brief, isolated monocytes using EasySep mouse monocyte isolation kit (Stemcell Technologies) from the spleen of treated and untreated EAE with 2DG were plated (25K/well) in a 96-well plate in RPMI media containing 10 mM glucose, 2 mM glutamine and 5% dialyzed fetal bovine serum. After 24 h, culture media was processed for the measurement of glucose and lactate using their respective assay kits from Promega. Total cellular levels of ATP were measured using ATPlite luminescence assay kit from Perkin-Elmer. Ten millimolar of the ATP solution was used to set up an ATP standard curve, and all samples were within the linear range of the assay.
Glucose glo and lactate glo assay kits
Manufactured by Promega
The Glucose Glo and Lactate Glo assay kits are laboratory products designed to measure glucose and lactate levels, respectively. The kits provide a quantitative and luminescent-based detection system for these analytes.
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2 protocols using glucose glo and lactate glo assay kits
1
Quantifying Metabolic Changes in EAE
2
Metabolic Profiling of Tumor Tissues
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Glucose and lactate metabolites were measured in tumor tissue collected from H446 and DMS79 tumors after 28 days using Glucose-Glo and Lactate-Glo assay kits (Promega). In brief, 3 mg of flash frozen tissue was homogenized in 50 mM Tris buffer containing 8:1 0.6N HCl. After homogenization, 8:1 1M Tris buffer was added to the tissue homogenates. Samples were assayed per manufacturer’s instruction and normalized to protein expression determined by DC Protein Assay Reagent (Bio-Rad).
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