Name: 2-NBDG
Brand: GlpBio
SKU: GC10289
Availability: InStock
Rating: 5 (39 reviews)

Product has been cited by 20 publications

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Related Biological Data

KT-NE reactivates dysfunctional TIDCs. (E) Representative fluorescent images and (F) quantification of GLUT1-based glucose uptake via 2-NBDG staining.
To verify the ability to transport glucose into (TI)DCs, 20 μM 2-NBDG (GlpBio) was utilized to treat DCs for 20 min at 37°C in the dark.
Nano Today 43 (2022): 101416. IF: 18.9615
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SR@PFeXCT alleviates aortic valve calcification in mice. a Experimental outline.
After overnight incubation, hVICs were incubated with glucose-free DMEM for 1 h, followed by 20min incubation in (2-(N-(7-Nitrobenz-2-oxa-1,3-diazol-4-yl)Amino)-2-Deoxyglucose)(2-NBDG, 100 μM, cat #GC10289, GlpBio) at 37 °C.
Nature Communications, 2024, 15(1): 557. PMID: 38228638 IF: 16.6009
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D)Fold change in protein expression of downstream p53 regulated pathway proteins in LNCaP L2A O/E + Enza cells from supplementary figure 3C (**p=≤0.01; *p=≤0.05; student t-test with Benjamini-Hochberg correction).
For glucose uptake assays, 2-[N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl) amino]-2-deoxy-D-glucose (2-NBDG) (GLPBIO) was added to the media of cells inculture at a final concentration of 50 μM for 15min in the cell culture incubators.
Oncogene (2023): 1-11. IF: 8.7558
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PDK4 promotes the calcification and glycolysis in VSMCs.k Glucose uptake was measured using a fluorescent analogue of glucose, 2-NBDG.
cells were plated onto coverslips and incubated with DMEM containing 10 μM 2-NBDG at room temperature for 1 h.
Cell death & disease 11.11 (2020): 1-23. PMID: 33203874 IF: 8.462
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AMPK inhibition blunted the activation effects of TPX on glucose uptake, the translocation of membrane GLUT4, and glycogen synthesis in the HG-induced IR cells. (A) The 2-NBDG assay was performed to examine glucose uptake activity with or without compound C (10 μM) administration.
The cells in each well were treated with 50 μM 2-NBDG(Glpbio) and 100 nM insulin for 30 min at 37°C.
Phytomedicine 114 (2023): 154740. PMID: 36965373 IF: 7.8996
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CD19+ B cells showed mitochondrial dysfunction at baseline and after T cell-dependent activation in D-LC. Representative flow cytometry graphs and bar charts of 2NBDG uptake (HC, n = 6; D-LC, n = 4)(A).
2NBDG(Glpbio) (working concentration 25 mg/ml, incubating at 37°C for 30 min) was used to assess glucose uptake by B cells.
Frontiers in Immunology 12 (2021). PMID: 34248941 IF: 7.562
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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.
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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.

Product Documents

Quality Control & SDS

View current batch:

Purity = 99.90%

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.

2-NBDGは、細胞のグルコース代謝を評価するためのトレーサーとして有用な、蛍光標識された2-デオキシグルコースの類似体である（Ex/Em: 475/550 nm）

2-NBDGは、細胞のグルコース取り込みを調べるための蛍光ラベル付きのグルコーストレーサーである。この分子は、グルコーストランスポーター（GLUT）によって細胞内に取り込まれる。細胞内で2-NBDGはヘキソキナーゼによってC-6位置でリン酸化され、その後細胞内に閉じ込められる。2-NBDGの取り込みは細胞の代謝活性や状態を評価する上で重要な指標であり、病態条件に応じて変化するため、疾患の進行状況を示す重要な指標となる。

2-NBDGはHepG2人間の肝癌細胞、L6ラットの骨格筋細胞、MCF-7乳がん上皮細胞やアストロサイトなど、多くの種類の細胞で試験管内で使用される。また、病態モデルやてんかんラット、高血糖、糖尿病、あるいはマウスのがんのゼノグラフトモデルなどでも使用される。

細胞内での2-NBDGの取り込みは、流式細胞計測法を用いて細胞が生きている状態で行われ、細胞が生成する蛍光を検出することによって行われる。また、蛍光マイクロプレートアッセイを使用して2-NBDGの細胞内濃度を測定することもできる。さらに、蛍光顕微鏡やCCDカメラを使用して簡単に2-NBDGを検出することができる。

2-NBDGは、グルコースと同じグルコーストランスポーター（GLUT）を介して細胞内に取り込まれるが、その後の代謝が異なる。2-NBDGはC-6位置でリン酸化され、2-NBDG-6-リン酸となり、これが細胞内によく保持される。そのため、2-DGやFDGなどの他のグルコーストレーサーと比較して、2-NBDGは高い時間分解能と空間分解能でシングルセルレベルでのin situ測定を可能にする。

References:
[1]. Zou C, Wang Y, Shen Z. 2-NBDG as a fluorescent indicator for direct glucose uptake measurement[J]. Journal of biochemical and biophysical methods, 2005, 64(3): 207-215.
[2]. O’Neil R G, Wu L, Mullani N. Uptake of a fluorescent deoxyglucose analog (2-NBDG) in tumor cells[J]. Molecular Imaging and Biology, 2005, 7(6): 388-392.
[3]. Tsytsarev V, Maslov K I, Yao J, et al. In vivo imaging of epileptic activity using 2-NBDG, a fluorescent deoxyglucose analog[J]. Journal of neuroscience methods, 2012, 203(1): 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 K I, Yao J, et al. In vivo imaging of epileptic activity using 2-NBDG, a fluorescent deoxyglucose analog[J]. Journal of neuroscience methods, 2012, 203(1): 136-140.

Cas No.	186689-07-6	SDF
同義語	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
溶解度	≥ 17.1mg/mL in Water with ultrasonic	Storage	-20°C, protect from light
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

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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FAQs & Troubleshooting

Why is there no significant difference in fluorescence staining intensity among 2-NBDG treatments?

Culture medium of 2-NBDG contains high level of glucose. Use culture medium which contains no glucose.

Why is 2-NBDG fluorescence intensity weak despite using highe concentration?

Cells were not starved before using 2-NBDG. Cells should be starved for 0.5-1 hr.

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2-NBDG (Synonyms: NBDGlucose)

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Product has been cited by 20 publications

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Product Documents

Quality Control & SDS

Protocol

Complete Stock Solution Preparation Table

モルアリティ計算機

希釈計算機

In vivo Formulation Calculator (Clear solution)

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FAQs & Troubleshooting

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