線粒體膜電位（Mitochondrial Membrane Potential,MMP）是判定細胞健康程度、線粒體膜通透性和細胞凋亡的一個重要指標，MMP的喪失通常與細胞凋亡的早期階段有關。評估線粒體功能狀態的基于細胞的檢測方法正在成為闡明線粒體活動在藥物誘導毒性、細胞凋亡級聯以及其他細胞和生化過程中的作用的有用工具。

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

作用機制

該染料的基本化學結構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入線粒體，當膜電位增加時，它的顏色會從綠色可逆地變為橙色（雙發射電位探針）。這種光物理特性是由于在膜極化時可逆地形成J-聚集體，導致在490nm處激發時，發射光從～530nm轉移到590nm。因此，具有低膜電位的線粒體將積累低濃度的染料，并表現出綠色熒光，而更高極化的線粒體將表現出橙色的熒光。

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

產品特點

● 耐光雙發射染料，能夠根據線粒體膜電位狀態發出綠色或橙色熒光

● 比JC-1熒光染料靈敏10倍，并具有卓-越的水溶性

● 無需洗滌步驟

● 適用于化學/環境毒性篩選

● 適用于高通量應用

如需購買ENZO產品，或咨詢產品技術問題，請聯系ENZO Life Science代理商欣博盛生物

實驗示例

圖1. 用MITO-ID® Membrane Potential reagent對HeLa細胞的線粒體進行染色，并通過熒光顯微鏡進行觀察。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胞質（FITC通道）。

圖2. 對照組和實驗組細胞的流式細胞儀分析。未經處理的Jurkat細胞（左）與用1μM CCCP處理15分鐘的Juekat細胞（右），用MITO-ID® Membrane Potential reagent對細胞進行染色，并使用流式細胞儀檢測。

產品信息

部分產品引用文獻

1. Discovery of the 3-Amino-1,2,4-triazine-Based Library as Selective PDK1 Inhibitors with Therapeutic Potential in Highly Aggressive Pancreatic Ductal Adenocarcinoma: D. Carbone, et al.; Int. J. Mol. Sci. 24, 3679 (2023) 

2. Gene signature predicting recurrence in oral squamous cell carcinoma is characterized by increased oxidative phosphorylation: J.K. Noh, et al.; Mol. Oncol. 17, 134 (2023) 

3. Cu(I) and Cu(II) Complexes Based on Lonidamine-Conjugated Ligands Designed to Promote Synergistic Antitumor Effects: F.D. Bello, et al.; Inorg. Chem. 61, 4919 (2022) 

4. Mitochondrial membrane potential-enriched CHO host: a novel and powerful tool for improving biomanufacturing capability: L. Chakrabarti, et al.; MAbs 14, 2020081 (2022)

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) 

7. Role of caspase-8 and/or-9 as biomarkers that can distinguish the potential to cause toxic-and immune related-adverse event, for the progress of acetaminophen-induced liver injury: T. Noda, et al.; Life Sci. 294, 120351 (2022), Application(s): Microplate reader 

8. Unveiling the Potential of Innovative Gold(I) and Silver(I) Selenourea Complexes as Anticancer Agents Targeting TrxR and Cellular Redox Homeostasis: M.D. Franco, et al.; Chemistry 28, e202201898 (2022) 

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

13. Depletion of mitochondrial components from extracellular vesicles secreted from astrocytes in a mouse model of fragile X syndrome: B.G. Ha, et al.; Int. J. Mol. Sci. 22, 410 (2021), Application(s): Fluorescence microscopy

14. Inhibiting autophagy targets human leukemic stem cells and hypoxic AML blasts by disrupting mitochondrial homeostasis: K.M. Dykstra, et al.; Blood Adv. 5, 2087 (2021)

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