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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
如需購(gòu)買ENZO產(chǎn)品,或咨詢產(chǎn)品技術(shù)問題,請(qǐng)聯(lián)系ENZO Life Science代理商欣博盛生物
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
產(chǎn)品貨號(hào) | ENZ-51018-0025/ ENZ-51018-K100 |
產(chǎn)品名稱 | MITO-ID® Membrane potential detection kit |
規(guī)格 | 1*25tests/1*100tests |
短期保存 | -20°C |
長(zhǎng)期保存 | -80°C |
試劑盒組分 | MITO-ID® MP Detection Reagent Necrosis Detection Reagent CCCP Control 10X Assay Buffer 1 50X Assay Buffer 2 |
應(yīng)用 | Flow Cytometry, Fluorescence microscopy, Fluorescent detection, HTS |
部分產(chǎn)品引用文獻(xiàn)
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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)
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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
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
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