2-NBDG (solution) 

2-NBDG (solution) 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.
Solvent and Concentration: Sterile PBS: 5 mM

1. 2-NBDG 工作液准备
1.1 工作液的配制
用预热好的无血清细胞培养基或 PBS 稀释储存液，可根据实际情况稀释相应的储备液。注意，如果溶剂为 DMSO，则必须考虑 DMSO 的细胞毒性，并应制备溶剂对照；如果溶剂为纯水，则工作液在加入细胞前需要过滤并除菌。
注：请根据实际情况调整 2-NBDG 工作液浓度，且现用现配。

2.细胞染色
2.1 悬浮细胞：离心收集细胞，加入 PBS 洗涤两次，每次 5 分钟。
贴壁细胞：弃去培养基，加入胰蛋白酶消化细胞。离心弃去上清后，加入 PBS 洗涤两次，每次 5 分钟。
2.2 加入 1 mL 2-NBDG 工作液，室温孵育 5-60 分钟。
2.3 400 g，4°C 离心 3-4 分钟，弃去上清。
2.4 加入 PBS 洗涤细胞两次，每次 5 分钟。
2.5 用 1 mL 无血清培养基或 PBS 重悬细胞后，使用流式细胞仪进行观察；
如果用荧光显微镜观察，可不消化直接染色；
如果进行活力测试，使用 ELISA 酶标仪记录 540/570 nm 的光密度 (O.D.)。细胞活力以对照比计算，并与药物对数浓度绘制曲线，计算 IC₅₀。

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

342.26

C₁₂H₁₄N₄O₈

186689-07-6

Room temperature in continental US; may vary elsewhere.

Please store the product under the recommended conditions in the Certificate of Analysis.

• [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]. 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]

  [3]. 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]

  [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]