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2-NBDG Venta (Synonyms: NBDGlucose)

Catalog No.GC10289

2-NBDG es un análogo de la 2-deoxi-glucosa marcado con fluorescencia útil como trazador para la evaluación del metabolismo celular de la glucosa (Ex/Em: 475/550 nm).

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2-NBDG Chemical Structure

Cas No.: 186689-07-6

Tamaño Precio Disponibilidad Cantidad
10mM (in 1mL Water)
165,00 $
Disponible
1mg
52,00 $
Disponible
5mg
101,00 $
Disponible
10mg
166,00 $
Disponible
500mg
2.756,00 $
Disponible
1g
4.568,00 $
Disponible

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Sample solution is provided at 25 µL, 10mM.

Product has been cited by 14 publications

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Protocol

Procedure for 2-NBDG uptake assay for MEFs [1]

1. Mouse embryonic fibroblasts (MEFs) are isolated from the embryos of C57BL/6 WT mouse (13.5 days).

2. Culture the MEF cells until reaching 80-90% confluence in 10 cm Petri dishes with DMEM growth medium in a humidified cell culture incubator (37 °C, 5% CO2).
Note: Don’t use MEFs beyond passage 3. MEFs usually become senescent at about passage 4 to 5.

3. Remove culture medium and wash cells one time with 10 ml 1x PBS.

4. Trypsinize cells using 4 ml of 0.05% trypsin-EDTA for 3 min at 37 °C.

Note: Use room temperature or pre-warmed 1x PBS from Step A3 to Step A9. Using chilled 1x PBS after Step A9.

5. Transfer cells to 15 ml polystyrene centrifuge tubes.

6. Harvest cells at 200 x g for 5 min by centrifugation.

7. Wash pelleted cells one time with 5 ml 1x PBS.

8. Count cells using a hemocytometer chamber.

9. Incubate 1 x 106 MEF cells in a 37 °C water bath for 2 h with 1 ml of PBS containing 100 μM 2-NBDG. Incubate the same number of MEFs in the water bath with 1 ml PBS without 2-NBDG as a negative control.

10. Pellet the cells at 200 x g for 5 min by centrifugation. After washing the cells with chilled 1x PBS, the cells are pelleted at 200 x g for 5 min by centrifugation.

11. Resuspend cells in 0.5 ml of ice-cold 1x PBS with 2% FBS.
Note: Always keep cells on ice after this step.

12. Filter cells through a 35 µm nylon mesh (the cell-strainer cap of the 5 ml round-bottom polystyrene tubes) to obtain a uniform single-cell suspension in a 5 ml tube.

13. Keep the samples on ice until analysis on a flow cytometer.

14. Perform flow cytometric analysis. Acquire 10,000 single-cell events per reaction.

15. Analyze fluorescence intensity

Procedure for 2-NBDG uptake assay for breast cancer cells [1]

Using the same procedure as MEFs’ uptake assay except incubating 1 x 106 MCF7 cells in a 37 °C water bath only for 30 min (instead of two hours for MEFs) with 1 ml of PBS containing 100 μM 2-NBDG.

10 mM stock of 2-NBDG: Dissolve 5 mg 2-NBDG in 1.46 ml PBS. Store at -20 °C in the dark

This protocol only provides a guideline, and should be modified according to your specific needs

References:
[1]. Dong, S., Baranwal, S., Garcia, A., Serrano-Gomez, S. J., Eastlack, S., Iwakuma, T., Mercante, D., Mauvais-Jarvis, F. and Alahari, S. K. (2017). Nischarin inhibition alters energy metabolism by activating AMP-activated protein kinase. J Biol Chem 292(41): 16833-16846.

Background

2-NBDG es un análogo de la 2-deoxi-glucosa marcado con fluorescencia útil como trazador para la evaluación del metabolismo celular de la glucosa (Ex/Em: 475/550 nm).

excitation and emission of 2-NBDG

La glucosa es una fuente necesaria de energía para mantener las actividades celulares y la homeostasis en los tejidos. El metabolismo de la glucosa es un objetivo importante en muchas enfermedades y cambia con la condición patológica, por lo tanto, la evaluación del metabolismo de la glucosa puede ser una indicación significativa en el progreso de las enfermedades.

2-NBDG se puede utilizar en muchos tipos de células in vitro, como las células HepG2 del hepatocarcinoma humano, las células musculares esqueléticas de rata L6, las células epiteliales del cáncer de mama MCF-7 y los astrocitos. También se utiliza en modelos de enfermedades como la epilepsia en ratas, la hiperglucemia, la diabetes o el modelo xenoinjerto de cáncer en ratones.

2-NBDG entra en las células a través de los transportadores de glucosa y posteriormente es fosforilado por la hexoquinasa y atrapado dentro de las células. La detección citométrica de flujo de la fluorescencia producida por las células se puede realizar para examinar la captación de 2-NBDG en células vivas, y la concentración intracelular del 2-NBDG transportado se puede medir con un ensayo microplaca fluorescente. También se puede detectar fácilmente con una microscopía de imágenes fluorescentes o una cámara CCD.

2-NBDG es un trazador de glucosa fluorescentemente etiquetado que se transporta a las células por el mismo transportador de glucosa (GLUT) que la glucosa. Una vez que 2-NBDG es absorbido por las células, se fosforila en la posición C-6 para dar 2-NBDG-6-fosfato, que se retiene bien en la célula. En comparación con otros trazadores de glucosa como 2-DG o FDG, 2-NBDG permite la medición in situ de 2-NBDG con alta resolución temporal y espacial a nivel celular individual. (adecuado para detección mediante microscopía de fluorescencia y citometría de flujo)

Principle of 2-NBDG

Justificación del ensayo de captación de glucosa 2-NBDG en células: Una vez que las células absorben el 2-NBDG, este se fosforila en la posición C-6 para generar 2-NBDG-6-fosfato en su metabolismo, el cual es bien retenido dentro de la célula. La intensidad de fluorescencia es proporcional a la actividad celular de captación de glucosa.

Referencias:
[1]. Zou C, Wang Y, Shen Z. 2-NBDG como indicador fluorescente para la medición directa de la captación de glucosa[J]. Journal of biochemical and biophysical methods, 2005, 64(3): 207-215.
[2]. O’Neil R G, Wu L, Mullani N. Captación de un análogo fluorescente de desoxiglucosa (2-NBDG) en células tumorales[J]. Molecular Imaging and Biology, 2005, 7(6): 388-392.
[3]. Tsytsarev V, Maslov K I, Yao J et al. Imágenes in vivo de actividad epiléptica utilizando el análogo fluorescente de desoxiglucosa 2-NBDG[J]. Journal of neuroscience methods ,2012;203:136–140.
[4] Yan Chen,Junjian Zhang,Xiang-yang Zhang,"2-NBDG as a Marker for Detecting Glucose Uptake in Reactive Astrocytes Exposed to Oxygen-Glucose Deprivation In Vitro".J Mol Neurosci (2015)55:126–130
[5] Tsytsarev V,Maslov KI,Yao J et al.In vivo imaging of epileptic activity using the fluorescent glucose analogs."Journal of Neuroscience Methods",2012;203:136-140

Chemical Properties

Cas No. 186689-07-6 SDF
Sinónimos NBDGlucose
Chemical Name (3R,4R,5S,6R)-6-(hydroxymethyl)-3-((7-nitrobenzo[c][1,2,5]oxadiazol-4-yl)amino)tetrahydro-2H-pyran-2,4,5-triol
Canonical SMILES OC[C@](O1)([H])[C@](O)([H])[C@@](O)([H])[C@](NC2=CC=C(N(=O)=O)C3=NON=C23)([H])C1([H])O
Formula C12H14N4O8 M.Wt 342.26
Solubility ≥ 17.1mg/mL in Water with ultrasonic Storage Store at -20°C
General tips Please select the appropriate solvent to prepare the stock solution according to the solubility of the product in different solvents; once the solution is prepared, please store it in separate packages to avoid product failure caused by repeated freezing and thawing.Storage method and period of the stock solution: When stored at -80°C, please use it within 6 months; when stored at -20°C, please use it within 1 month.
To increase solubility, heat the tube to 37°C and then oscillate in an ultrasonic bath for some time.
Shipping Condition Evaluation sample solution: shipped with blue ice. All other sizes available: with RT, or with Blue Ice upon request.

Complete Stock Solution Preparation Table

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1 mg 5 mg 10 mg
1 mM 2.9218 mL 14.6088 mL 29.2176 mL
5 mM 0.5844 mL 2.9218 mL 5.8435 mL
10 mM 0.2922 mL 1.4609 mL 2.9218 mL
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Related Video

    Glucose Uptake Assay with 2-NBDG

    2-NBDG- GlpBio

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