DiBAC4(5) 膜電位熒光探針

產品簡介：

慢速響應探針（Slow-response probes）包括陽離子碳花菁、羅丹明和陰離子氧鎓醇，都適用于檢測非興奮細胞因呼吸活性、離子通道滲透性、藥物結合和其他因素引起的平均膜電位變化。

DiBAC4(5)(Bis-(1,3-dibutylbarbituric acid)pentamethine oxonol)是帶陰離子的慢速響應電位敏感探針，在pH4.5~7.5的范圍內表現(xiàn)出非pH敏感的熒光特征（最大激發(fā)和發(fā)射波長分別為590nm和616nm）。能夠進入去極化細胞，之后結合到胞內蛋白或膜上。一旦結合，DiBAC4(5)的熒光特性加強，往紅光區(qū)遷移。增加的細胞去極化引起增多的染料內流和增高的熒光信號。DiBAC4(5)的負電荷阻止其進入線粒體，因此，適用于細胞膜電位測定。

產品特性：

1） CAS NO：63560-89-4

2） 同義名：Bis-(1,3-dibutylbarbituric acid)pentamethine oxonol

3） 分子式：C29H42N4O6 

4） 分子量：542.67g/mol

5） Ex/Em：590/616 nm

6） 溶解性：溶于DMSO

保存條件: 

-20℃干燥避光保存，有效期至少一年。

產品使用：（僅供參考）

文獻來源：Zhang Y et al. A Membrane Potential- and Calpain-Dependent Reversal of Caspase-1 Inhibition Regulates Canonical NLRP3 Inflammasome.Cell Rep. 2018 Aug 28;24(9):2356-2369.e5.

實驗方法（膜電位染色）：DiBAC4(5) is a translational MP dyes that redistributes within the cell membrane as MP changes. MP depolarization results in an increase in fluorescence intensity of cells at Ex/Em = 590/616nm. Conversely, hyperpolarization is indicated by a decrease in fluorescence. PMA-differentiated THP-1 cells were cultured in 35mm imaging dish with a concentration of 4x105cells/ml. Cells were stained in 200 nM DiBAC4(5)in 5mM K+ buffer at 37°C for 30 min. Then the staining buffer was removed without washing and crystals (silica, Al(OH)3, CPPD) or inhibitors (glyburide or CFTRinh-172) in fresh 5mM K+buffer containing 200nM DiBAC4(5) was added to the cells. For high K+ condition, staining buffer was replaced with 100mM K+ buffer containing 200nM DiBAC4(5). We captured 5 different fields at each time-point on fluorescent microscope, and analyzed the fluorescence intensity of all the cells by ImageJ.

實驗結果（膜電位染色）：

Fig. Depolarization or Hyperpolarization of Membrane Potential May Affect the Activation of NLRP3 Inflammasome.THP-1 cells were stained with DiBAC4(5) to check the effect of CFTRinh-172 on MP. Representative images and relative fluorescence unit (RFU) plotting are shown. Each point was mean ± SEM of cells from at least 12 fields in 2 independent experiments. Scale bar, 20 μm.

注意事項：

1.      本品用有機溶劑（比如：DMSO）配制的儲存液，請根據(jù)單次用量分裝，置于≤-20℃避光保存。

2.      熒光染料均存在淬滅問題，請盡量注意避光，以減緩熒光淬滅。

3.      為了您的安全和健康，請穿實驗服并戴一次性手套操作。

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