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.

Guide (Following is our recommended protocol. This protocol only provides a guideline, and should be modified according to your specific needs).
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.
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 Cell preparation.
For suspension cells: Centrifuge at 1,000 g at 4°C 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°C 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°C for 3-4 minutes and then discard the supernatant.
2.4 Wash twice with PBS, 5 minutes each time.
2.5 Resuspend cells with 1 mL serum-free cell culture medium or PBS, and then detect by fluorescence microscope or flow cytometer.
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.

MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.

342.26

C₁₂H₁₄N₄O₈

542

488

186689-07-6

Solid

Light yellow to orange

O=C[C@H](NC1=CC=C([N+]([O-])=O)C2=NON=C21)[C@H]([C@@H]([C@@H](CO)O)O)O

Room temperature in continental US; may vary elsewhere.

-20°C, protect from light

*In solvent : -80°C, 6 months; -20°C, 1 month (protect from light)

* Please refer to the solubility information to select the appropriate solvent. Once prepared, please aliquot and store the solution to prevent product inactivation from repeated freeze-thaw cycles.
Storage method and period of stock solution: -80°C, 6 months; -20°C, 1 month (protect from light). When stored at -80°C, please use it within 6 months. When stored at -20°C, please use it within 1 month.

* Note: If you choose water as the stock solution, please dilute it to the working solution, then filter and sterilize it with a 0.22 μm filter before use.

• [1]. Yamada K, et al. A real-time method of imaging glucose uptake in single, living mammalian cells. Nat Protoc. 2007;2(3):753-62.  [Content Brief]

  [2]. Katsuya Yamada, et al. A real-time method of imaging glucose uptake in single, living mammalian cells. Nat Protoc. 2007;2(3):753-62.  [Content Brief]

  [3]. Zou C, et al. 2-NBDG as a fluorescent indicator for direct glucose uptake measurement. J Biochem Biophys Methods. 2005 Sep 30;64(3):207-15.  [Content Brief]

  [4]. Zou C, et al. 2-NBDG as a fluorescent indicator for direct glucose uptake measurement. J Biochem Biophys Methods. 2005 Sep 30;64(3):207-15.  [Content Brief]