2-NBDG (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).
Products are for research use only. Not for human use. We do not sell to patients.
Cas No.: 186689-07-6
Sample solution is provided at 25 µL, 10mM.
- Ecotox Environ Safe 243 (2022): 113996.PMID:36030680
- Bioorganic Chemistry (2023): 106341.
- Oncogene (2023): 1-11.
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- Front Pharmacol 13 (2022): 875014.PMID:35694255
- Ind Eng Chem Res (2023).
- Phytomedicine 114 (2023): 154740.PMID:36965373
- Nat Commun 15.1 (2024):557.PMID:38228638
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Related Biological Data
Representative fluorescent images of GLUT1-based glucose uptake via 2-NBDG staining.
To verify the ability to transport glucose into (TI)DCs, 20 μM 2-NBDG was utilized to treat DCsfor 20 min at 37 °C in the dark. Hoechst 33342 (10 μug/mL) was used to visualize the nucleus. Corresponding fluorescent images were captured by high resolution fluorescence microscopy (F900, Edinburgh Instruments Ltd., UK).
Nano Today 43 (2022): 101416. -
Related Biological Data
Glucose uptake was measured using a fluorescent analogue of glucose, 2-NBDG. Scale bars: 10 μm, N = 40- -50 cells per group.
Glucose uptake ability of VSMCs was evaluated by using the fluores-cent glucose 2-NBDG (GlpBio, USA) according to the manufactur-er’s instruction. cells were plated onto coverslips and incubated with DMEM containing 10 μM 2-NBDG at room temperature for 1 h.Cell Death Dis.
2020 Nov 17;11(11):991. -
Related Biological Data
Effect of CrEL on glucose transport.2-NBDG is a fluorescent glu-cose analog. INK-128 (mTORi) was used as a control for de-creased glucose transport.
Culture medium was then removed from each well, and treated with medium with or without 200 μM 2-NBDG (GlpBio) for 20 minutes.
iScience.2021 Sep 25;24(10):103170.
Quality Control & SDS
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Purity = 99.90%
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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.
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).
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)
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
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. |
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Shipping Condition | Evaluation sample solution: shipped with blue ice. All other sizes available: with RT, or with Blue Ice upon request. |
Prepare stock solution | |||
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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Method for preparing DMSO master liquid: mg drug pre-dissolved in μL DMSO ( Master liquid concentration mg/mL, Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug. )
Method for preparing in vivo formulation: Take μL DMSO master liquid, next addμL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O, mix and clarify.
Method for preparing in vivo formulation: Take μL DMSO master liquid, next add μL Corn oil, mix and clarify.
Note: 1. Please make sure the liquid is clear before adding the next solvent.
2. Be sure to add the solvent(s) in order. You must ensure that the solution obtained, in the previous addition, is a clear solution before proceeding to add the next solvent. Physical methods such as vortex, ultrasound or hot water bath can be used to aid dissolving.
3. All of the above co-solvents are available for purchase on the GlpBio website.
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