| Assay Method Information | |
| | [3H]-2-Deoxy-D-glucose ([3H]-2-DG) Uptake Assay |
| Description: | The stably transfected CHO cells were seeded in poly 24-well plates and were grown to 80−90% confluence. Cells were rinsed with PBS buffer and incubated in 280 μL of PBS buffer containing 61.73 μM [3H]-2-DG (PerkinElmer, Boston, MA) in the presence and absence of 50 μM test compound for 5 min. The known hGLUT1 inhibitor phloretin (Accela ChemBio, San Diego, CA) was included as a positive control. Nonradiolabeled 2-DG (40 mM) (Sigma, St. Louis, MO) was included inorder to calculate specific inhibition of predicted inhibitors. Compounds were obtained from the National Cancer Institute (NCI) or purchased from commercial vendors. The reaction was terminated by washing cells twice with ice-cold PBS, followed by the addition of 200 μL of lysis buffer (0.1% (w/v) SDS, 0.1 N NaOH).Intracellular radioactivity was determined by scintillation counting. |
| Affinity data for this assay | |
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