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2-NBDG Sale

(Synonyms: 2-(N-7-硝基-2,1,3-苯并恶二唑-4-氨基)-2-脱氧-D-葡萄糖) 目录号 : GC10289

2-NBDG是一种荧光标记的2-脱氧葡萄糖类似物,可用作细胞葡萄糖代谢评估的示踪剂(激发/发射波长:475/550纳米)。

2-NBDG Chemical Structure

Cas No.:186689-07-6

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10mM (in 1mL Water)
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Sample solution is provided at 25 µL, 10mM.

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Description

2-NBDG is a fluorescence-labeled 2-deoxy-glucose analog useful as a tracer for evaluation of cellular glucose metabolism (Ex/Em: 475/550 nm).

excitation and emission of 2-NBDG

Glucose is a necessary source of energy for sustaining cell activities and homeostasis in tissues. Glucose metabolism is an important target in many diseases and changed with the pathological condition, therefore, evaluation of glucose metabolism can be a significant indication in disease progressions.

2-NBDG can be used in many kinds of cells in vitro, such as HepG2 human hepatocarcinoma cells, L6 rat skeletal muscle cells, MCF-7 breast cancer epithelial cells and astrocytes, it is also used in disease models, epilepsy rat, hyperglycemia, diabetes or mouse xenograft model of cancer.

2-NBDG enters cells through glucose transporters and is subsequently phosphorylated by hexokinase and trapped inside cells. Flow cytometric detection of fluorescence produced by cells can be performed to examine 2-NBDG uptake into living cells, and the intracellular concentration of transported 2-NBDG can be measured with a fluorescence microplate assay. It can be detected with a fluorescence imaging microscopy or CCD camera simply as well.

2-NBDG is a fluorescently labeled glucose tracer that is transported into cells by the same glucose transporter (GLUT) as glucose. Once 2-NBDG is taken up by cells, it is phosphorylated at the C-6 position to give 2-NBDG-6-phosphate, which is well retained in the cell. Compared to other glucose tracers such as 2-DG or FDG, 2-NBDG enables in situ measurement of 2-NBDG with high temporal and spatial resolution at the single-cell level. (suitable for fluorescence microscopy and flow cytometry detection)

Principle of 2-NBDG

Rationale for 2-NBDG glucose uptake assay in cells: Once 2 NBDG is taken up by cells, it is phosphorylated at the C-6 position to generate 2-NBDG-6-phosphate in 2 NBDG metabolism, which is well retained in the cell, the fluorescence intensity is proportional to the cellular glucose uptake activity.

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.

2-NBDG是一种荧光标记的2-脱氧葡萄糖类似物,可用作细胞葡萄糖代谢评估的示踪剂(激发/发射波长:475/550纳米)。

excitation and emission of 2-NBDG

葡萄糖是维持组织细胞活动和稳态所必需的能量来源。葡萄糖代谢在许多疾病中都是一个重要的靶点,并且会随着病理情况而改变,因此评估葡萄糖代谢可以成为一种重要的指标来判断疾病进展情况。

2-NBDG可以在许多种体外细胞中使用,例如HepG2人类肝癌细胞、L6大鼠骨骼肌细胞、MCF-7乳腺癌上皮细胞和星形胶质细胞,它还用于疾病模型,如癫痫大鼠、高血糖、糖尿病或小鼠异种移植的癌症模型。

2-NBDG通过葡萄糖转运体进入细胞,并被己糖激酶磷酸化并困在细胞内。可以使用流式细胞术检测细胞产生的荧光来检查2-NBDG进入活细胞的情况,也可以使用荧光微孔板分析法测量转运的2-NBDG在细胞内的浓度。它还可以简单地用荧光成像显微镜或CCD相机进行检测。

2-NBDG是一种荧光标记的葡萄糖示踪剂,通过与葡萄糖相同的葡萄糖转运体(GLUT)进入细胞。一旦2-NBDG被细胞吸收,它会在C-6位置磷酸化成为2-NBDG-6-phosphate,并且能够很好地保留在细胞内。与其他葡萄糖示踪剂如2-DG或FDG相比,2-NBDG可以以高时间和空间分辨率在单个细胞水平上原位测量2-NBDG。(适用于荧光显微镜和流式细胞术检测)

Principle of 2-NBDG

2-NBDG葡萄糖摄取实验在细胞中的原理:一旦细胞吸收了2-NBDG,它会在C-6位置被磷酸化,生成2-NBDG-6-磷酸盐,在2 NBDG代谢中很好地保留在细胞内,荧光强度与细胞内葡萄糖摄取活性成正比。

实验参考方法

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.

化学性质

Cas No. 186689-07-6 SDF
别名 2-(N-7-硝基-2,1,3-苯并恶二唑-4-氨基)-2-脱氧-D-葡萄糖
化学名 (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
分子式 C12H14N4O8 分子量 342.26
溶解度 ≥ 17.1mg/mL in Water with ultrasonic 储存条件 -20°C, protect from light
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