2-NBDG

2-NBDG is a fluorescently-labeled deoxyglucose analog that is used primarily to directly monitor glucose uptake by living cells and tissues. It is also used as a topical contrast reagent for the detection of neoplasia. 2-NBDG can be used in real-time confocal, high-resolution, or wide-field fluorescence microscopy as well as in flow cytometry. The probe can be excited by the Argon laser at 488 nm to give the environment-sensitive fluorescence. It has lower photostability than the rhodamine-based fluorescent probes. In Vitro:Preparation of 2-NBDG working solution
1.1 Preparation of the stock solution
Dissolve 1 mg of 2-NBDG in 2.92 mL of DDH₂O to obtain 1 mM of 2-NBDG.
Note: It is recommended to store the stock solution at -20 ℃ or -80 ℃ away from light and avoid repetitive freeze-thaw cycles.
1.2 Preparation of 2-NBDG working solution.
Dilute the stock solution in serum-free cell culture medium or PBS to obtain 10-200 μM of 2-NBDG working solution.
Note: Please adjust the concentration of 2-NBDG working solution according to the actual situation.
Cell staining
2.1 For suspension cells: Centrifuge at 1,000 g at 4℃ for 3-5 minutes and then discard the supernatant. Wash twice with PBS, 5 minutes each time.
For adherent cells: Discard the cell culture medium, and add trypsin to dissociate cells to make a single-cell suspension. Centrifuge at 1000 g at 4℃ for 3-5 minutes and then discard the supernatant. Wash twice with PBS, 5 minutes each time.
2.2 Add 1 mL of 2-NBDG working solution, and then incubate at room temperature for 5-60 minutes.
2.3 Centrifuge at 400 g at 4℃ for 3-4 minutes and then discard the supernatant.
2.4 Wash twice with PBS, 5 minutes each time.
2.5 Resuspend cells with serum-free cell culture medium or PBS. .If test viability, recorded the optical density (O.D.) at 540/570 nm. Cell viability was calculated as a control ratio and plotted against the logarithmic concentration of the drug to calculate IC₅₀.