線粒體膜電位（Mitochondrial Membrane Potential,MMP）是判定細(xì)胞健康程度、線粒體膜通透性和細(xì)胞凋亡的一個(gè)重要指標(biāo)，MMP的喪失通常與細(xì)胞凋亡的早期階段有關(guān)。評(píng)估線粒體功能狀態(tài)的基于細(xì)胞的檢測(cè)方法正在成為闡明線粒體活動(dòng)在藥物誘導(dǎo)毒性、細(xì)胞凋亡級(jí)聯(lián)以及其他細(xì)胞和生化過(guò)程中的作用的有用工具。

Enzo Life Sciences的MITO-ID® Membrane potential detection kit包含一種雙發(fā)射陽(yáng)離子染料，用于檢測(cè)活細(xì)胞中的線粒體膜電位（MMP）。在有能量或活躍的細(xì)胞中，MITO-ID® 膜電位試劑因其相對(duì)負(fù)電荷而在線粒體中迅速聚集成發(fā)橙色熒光的聚合體，而在細(xì)胞質(zhì)中則以發(fā)綠色熒光的單體存在。然而，在MMP受損的細(xì)胞中，MITO-ID®膜電位試劑主要以綠色熒光單體存在于整個(gè)細(xì)胞質(zhì)中，在線粒體中不再表現(xiàn)出橙色熒光。

作用機(jī)制

該染料的基本化學(xué)結(jié)構(gòu)由高度共軛的部分組成，使正電荷廣泛離域。該染料能夠選擇性地進(jìn)入線粒體，當(dāng)膜電位增加時(shí)，它的顏色會(huì)從綠色可逆地變?yōu)槌壬p發(fā)射電位探針）。這種光物理特性是由于在膜極化時(shí)可逆地形成J-聚集體，導(dǎo)致在490nm處激發(fā)時(shí)，發(fā)射光從～530nm轉(zhuǎn)移到590nm。因此，具有低膜電位的線粒體將積累低濃度的染料，并表現(xiàn)出綠色熒光，而更高極化的線粒體將表現(xiàn)出橙色的熒光。

 MITO-ID® Membrane potential detection kit線粒體膜電位檢測(cè)試劑盒

產(chǎn)品特點(diǎn)

● 耐光雙發(fā)射染料，能夠根據(jù)線粒體膜電位狀態(tài)發(fā)出綠色或橙色熒光

● 比JC-1熒光染料靈敏10倍，并具有卓-越的水溶性

● 無(wú)需洗滌步驟

● 適用于化學(xué)/環(huán)境毒性篩選

● 適用于高通量應(yīng)用

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實(shí)驗(yàn)示例

圖1. 用MITO-ID® Membrane Potential reagent對(duì)HeLa細(xì)胞的線粒體進(jìn)行染色，并通過(guò)熒光顯微鏡進(jìn)行觀察。橙色熒光聚集體定位于線粒體（橙色通道），而綠色熒光單體主要定位于細(xì)胞質(zhì)（FITC通道）。

圖2. 對(duì)照組和實(shí)驗(yàn)組細(xì)胞的流式細(xì)胞儀分析。未經(jīng)處理的Jurkat細(xì)胞（左）與用1μM CCCP處理15分鐘的Juekat細(xì)胞（右），用MITO-ID® Membrane Potential reagent對(duì)細(xì)胞進(jìn)行染色，并使用流式細(xì)胞儀檢測(cè)。

產(chǎn)品信息

部分產(chǎn)品引用文獻(xiàn)

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5. New glycoconjugation strategies for Ruthenium(II) arene complexes via phosphane ligands and assessment of their antiproliferative activity: D. Iacopini, et al.; Bioorg. Chem. 126, 105901 (2022)

6. New tricyclic systems as photosensitizers towards triple negative breast cancer cell: M. Barreca, et al.; Arch. Pharm. Res. 45, 806 (2022) 

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9. Altered inflammatory response in FMRP-deficient microglia: J.M. Parrott, et al.; iScience 24, 103293 (2021), Application(s): Fluorescence microscopy 

10. ApoE4 impairs neuron-astrocyte coupling of fatty acid metabolism: G. Qi, et al.; Cell. Rep. 34, 108572 (2021), Application(s): Microplate reader 

11. Botrytis cinerea methyl isocitrate lyase mediates oxidative stress tolerance and programmed cell death by modulating cellular succinate levels: L. Oren-Young, et al.; Fungal Genet. Biol. 146, 103484 (2021), Application(s): Conidia (fungus spore); microscopy 

12. Copper (II) complexes containing natural flavonoid pomiferin show considerable in vitro cytotoxicity and anti-inflammatory effects: J. Vanco, et al.; Int. J. Mol. Sci. 22, 7626 (2021), Application(s): Flow cytometry

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15. Selective striatal cell loss is ameliorated by regulated autophagy of the cortex: K. Cho & G.W. Kim; Life Sci. 282, 119822 (2021), Application(s): Flow cytometry