2-NBDG（2-NBD-Glucose，86689-07-6）是熒光標(biāo)記的脫氧葡萄糖類似物，用于直接監(jiān)測活細(xì)胞和組織對葡萄糖的攝取。 2-NBD-Glucose用作檢測瘤形成的局部造影劑。 攝取可以通過實時共聚焦、高分辨率或?qū)捯晥鰺晒怙@微鏡和流式細(xì)胞術(shù)進行監(jiān)測。

溶解度Solubility溶于乙醇至 50 mM，溶于 DMSO 至 25 mM性狀solid,固體粉末儲藏條件 Storage conditionsstorage at -4℃ (1-2weeks), longer storage period at -20℃ (1-2years)

1、2-NBDG儲存液制備方法？

向 1mg粉末中加 100μl 無水DMSO 或者 乙醇，進行充分溶解混勻，制備成10mg/ml的儲存液，按照單次用量進行分裝后，在-20oC避光條件下冷藏存儲。若已經(jīng)配制成水溶液后，建議存周期不超過一天。

如實驗需要延長保存周期，最好是以粉末的形式在-20oC條件下保存。先用乙醇溶解固體粉末，然后分裝成小量裝入獨立瓶內(nèi)，使用真空泵將溶劑抽干即可。此時不推薦用DMSO或DMF為溶劑，因揮發(fā)性比較差。

2、2-NBDG工作濃度？

2-NBDG 的常用工作濃度大約10-600μM。細(xì)菌或者酵母菌染色的建議濃度范為100-600μM，原代和傳代培養(yǎng)哺乳動物細(xì)胞建議工作濃度范圍為10-200μM。

日常實驗過程 2-NBDG?工作濃度范圍為： 30-300μM。

細(xì)胞需要在在合適的溫度條件下孵育，以充分檢測綠-黃色熒光。攝取能力強弱受溫度影響，孵育時間有幾秒鐘，到30min，甚至更長，取決于探針終濃度、細(xì)胞類型、培養(yǎng)條件和一些其他因素。

案例：

編輯搜圖

Delivery-corrected 2-NBDG uptake inversely correlates with blood glucose concentration. (A) Representative images show the kinetics of 2-NBDG uptake in vivo in non-tumor window chambers. The same mouse was given 6 mM or 10 mM 2-NBDG on subsequent days and imaged for 60 minutes following injection. (B) Averaged 2-NBDG kinetics for a cohort of mice injected with 0.1 mL of either 6 mM or 10 mM 2-NBDG. At 5 minutes post-injection (2-NBDG05), the fluorescence ratio of the dose groups (2-NBDG05,10 mM/2-NBDG05,6 mM) was proportional to molarity (p<0.01). The table shows the expected ratio of 10 mM/6 mM fluorescence, if all differences in fluorescence were due to dose. 2-NBDG05,10 mM/2-NBDG05,6 mM corresponds to the ratio of 10 mM and 6 mM fluorescence intensities at t = 5 min. The ratio RD,10 mM/RD,6 mM corresponds to the rate of 2-NBDG delivery for 10 mM and 6 mM. Each group in panel B contains the same n = 7 subjects. p values are from a student's paired t-test. Error bars show standard error. Values in table are mean ± standard error. (C) RD was strongly correlated with 2-NBDG fluorescence at 5 minutes (p<0.001). RD did not correlate with 2-NBDG60 (not shown). (D) 2-NBDG60/RD was inversely correlated with baseline blood glucose in normal mice (R = ?0.61, p = 0.02). 2-NBDG60 was also correlated with blood glucose (R = ?0.52, p = 0.05, not shown). For animals that received both 6 mM and 10 mM doses, the average values of the endpoints (2-NBDG05, 2-NBDG60, and 2-NBDG60/RD) for both doses were used in calculating the correlations. These subjects are denoted by “mean” in the legend. n = 15 mice for (C) and (D).