2-NBDG 是一种荧光标记的脱氧葡萄糖类似物，主要用于直接监测活细胞和组织的葡萄糖摄取。它还可用作局部对比剂，用于检测肿瘤。2-NBDG 可用于实时共焦、高分辨率或广场荧光显微镜以及流式细胞术。探针可以被 488 nm 的激光激发，产生环境敏感荧光。光稳定性低于罗丹明荧光探针。
| 生物活性 | 2-NBDG 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. |
体外研究
(In Vitro) | 1. 2-NBDG 工作液准备
1.1 制备储存液
用 2.92 mL DDH2O 稀释 1 mg 2-NBDG 配制 1 mM 储备液。
1.2 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，配制成 10-200 μM 的 2-NBDG 工作液。
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℃ 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，使用显微镜进行观察，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。
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| 分子量 | |
| 性状 | |
| Formula | |
| CAS 号 | |
| Emission (Em) | Em 495-570 nm Green |
| Excitation (Ex) | Ex 450-495 nm Blue |
| 运输条件 | Room temperature in continental US; may vary elsewhere. |
| 储存方式 | | Powder | -20°C | 3 years | | In solvent | -80°C | 6 months | | -20°C | 1 month |
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| 溶解性数据 | In Vitro: H2O : 5 mg/mL(14.61 mM;ultrasonic and warming and heat to 60℃) 配制储备液 | 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 |
*请根据产品在不同溶剂中的溶解度选择合适的溶剂配制储备液；一旦配成溶液，请分装保存，避免反复冻融造成的产品失效。
储备液的保存方式和期限：-80℃, 6 months; -20℃, 1 month。-80℃ 储存时，请在 6 个月内使用，-20℃ 储存时，请在 1 个月内使用。 In Vivo: 请根据您的实验动物和给药方式选择适当的溶解方案。以下溶解方案都请先按照In Vitro方式配制澄清的储备液，再依次添加助溶剂： ——为保证实验结果的可靠性，澄清的储备液可以根据储存条件，适当保存；体内实验的工作液，建议您现用现配，当天使用；
以下溶剂前显示的百
分比是指该溶剂在您配制终溶液中的体积占比；如在配制过程中出现沉淀、析出现象，可以通过加热和/或超声的方式助溶 1. 请依序添加每种溶剂： PBS Solubility: 3.33 mg/mL (9.73 mM); Clear solution; Need ultrasonic and warming and heat to 60℃
*以上所有助溶剂都可在本网站选购。 |
| 染色示例 | Description: 2-NBDG can be used as a fluorescent indicator for monitoring glucose uptake in living cells.Method: For 2-NBDG uptake assay.1. Incubate cells with 2-NBDG (50 μM; 30 min; 37℃; 5% CO2) in glucose-free DMEM.2. Wash cells three times with warm PBS and use ImageXpress Micro Confocal for image. Description: 2-NBDG can be used as a fluorescent indicator for monitoring glucose uptake in living cells.Method: For 2-NBDG uptake assay.1. Incubate cells with low-glucose DMEM containing 2-NBDG (150 μg/mL; 60 min; 37℃).2. Wash cells for five times with PBS and use Nikon a Eclipse Ti-s microscopy (Nikon, Tokyo, Japan) to observe fluorescence. Description: 2-NBDG can be used as a fluorescent indicator for monitoring glucose uptake in living cells.Method: For 2-NBDG uptake assay.1. Incubate cells in low-glucose DMEM with 2-NBDG (150 μg/mL; 60 min; 37℃).2. Wash cells five times with PBS and use a Nikon Eclipse Ti-s microscopy (Nikon) to observe fluorescence. Description: 2-NBDG can be used as a fluorescent indicator for monitoring glucose uptake in living cells.Method: For 2-NBDG uptake assay.1. Incubate cells with 2-NBDG (100 μM; 30 min).2. Thoroughly wash cells and the staining is observed by a fluorescence microscope with Cellsens Dimention 1.15 software (Olympus, Tokyo, Japan).
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