線粒體膜電位細(xì)胞毒性檢測(cè)試劑盒介紹

Enzo Life Sciences的MITO-ID® Membrane potential cytotoxicity kit利用陽(yáng)離子雙發(fā)射染料檢測(cè)線粒體膜電位（MMP）的波動(dòng)，該染料在細(xì)胞質(zhì)中以綠色熒光單體的形式存在，在線粒體中以橙色熒光J聚集體的形式聚集。具有低膜電位的線粒體將積累低濃度的染料并呈現(xiàn)綠色熒光，而更高度極化的線粒體將呈現(xiàn)橙紅色熒光。隨著線粒體功能受損加劇，細(xì)胞表現(xiàn)出從橙色熒光到綠色熒光的轉(zhuǎn)變。該試劑盒是一種獨(dú)-特的HTS測(cè)定法，無(wú)需洗滌或去除培養(yǎng)基即可實(shí)時(shí)監(jiān)測(cè)線粒體膜電位。

產(chǎn)品特點(diǎn)

● 靈敏度是JC-1熒光染料的10倍，并具有卓-越的水溶性

● 具備光穩(wěn)定性的雙發(fā)射染料

● 無(wú)需漂洗/換液步驟

● 單獨(dú)的MITO-ID®檢測(cè)可用于檢測(cè)線粒體質(zhì)量

● 可在較低的藥物/劑量濃度下檢測(cè)細(xì)胞毒性

● 沒(méi)有使用JC-1染料時(shí)出現(xiàn)的溶劑偽影

● 適用于高通量應(yīng)用

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實(shí)驗(yàn)示例

圖1. 檢測(cè)線粒體紊亂的靈敏度是JC-1的10倍。使用MITO-ID®膜電位染料（紅色）或JC-1（藍(lán)色）在經(jīng)CCCP處理的HeLa細(xì)胞中評(píng)估線粒體膜電位（MMP）。使用傳統(tǒng)的熒光酶標(biāo)儀檢測(cè)，結(jié)果顯示MMP 隨著CCCP濃度的增加而減少，橙色熒光減少。染料的水溶性優(yōu)化和無(wú)需洗滌的實(shí)驗(yàn)方案最大限度減小了可變性，因此Z因子（>0.9）高于使用JC-1.

圖2. 在藥物篩選中實(shí)時(shí)檢測(cè)有絲分裂毒性。使用BioTek Synergy™ Mx熒光酶標(biāo)儀對(duì)線粒體膜電位變化的時(shí)間進(jìn)程研究。HeLa細(xì)胞與MITO-ID®膜電位染料在室溫下孵育30分鐘（不去除血清或培養(yǎng)基）。添加魚(yú)藤酮分別達(dá)到1µM、3µM和9µM的濃度。橙色信號(hào)的減少證明染料對(duì)魚(yú)藤酮有反應(yīng)。

產(chǎn)品信息

部分產(chǎn)品引用文獻(xiàn)

1. Novel Silver Complexes Based on Phosphanes and Ester Derivatives of Bis(pyrazol-1-yl)acetate Ligands Targeting TrxR: New Promising Chemotherapeutic Tools Relevant to SCLC Management: M. Pellei, et al.; Int. J. Mol. Sci. 24, 4091 (2023)

2. FUNDC1 regulates receptor-mediated mitophagy independently of the PINK1/Parkin-dependent pathway in rotenone-treated SH-SY5Y cells: S.Y. Park, et al.; Food Chem. Toxicol. 137, 111163 (2020), Application(s): Fluorescence microscopy and microplate reader

3. Inhibitory role of TRIP-Br1/XIAP in necroptosis under nutrient/serum starvation: Z. Sandag, et al.; Mol. Cells 43, 236 (2020)

4. NAD hydrolysis by the tuberculosis necrotizing toxin induces lethal oxidative stress in macrophages: D. Pajuelo, et al.; Cell. Microbiol. 22, e13115 (2020), Application(s): THP-1 macrophages; microplate reader

5. Impaired autophagic and mitochondrial functions are partially restored by ERT in Gaucher and Fabry diseases: M.M. Ivanova, et al.; PLoS One 14, e0210617 (2019), Application(s): Fluorescence microscopy

6. Inhibitory role of AMPactivated protein kinase in necroptosis of HCT116 colon cancer cells with p53 null mutation under nutrient starvation: D.T. Le, et al.; Int. J. Oncol. 54, 702 (2019)

7. ROS as a novel indicator to predict anticancer drug efficacy: T. Zaidieh, et al.; BMC Cancer 19, 1224 (2019)

8. TREM1/3 deficiency impairs tissue repair after acute kidney injury and mitochondrial metabolic flexibility in tubular epithelial cells: A. Tammaro, et al.; Front. Immunol. 10, 1469 (2019), Application(s): Microplate reader

9. Anti-cancerous effect of cis-khellactone from Angelica amurensis through the induction of three programmed cell deaths: S. Jung, et al.; Oncotarget 9, 16744 (2018), Application(s): Microplate reader

10. Blue light phototoxicity toward human corneal and conjunctival epithelial cells in basal and hyperosmolar conditions: V. Marek, et al.; Free Radic. Biol. Med. 126, 27 (2018), Application(s): Microplate reader

11. cGAS drives noncanonical-inflammasome activation in age-related macular degeneration: N. Kerur, et al.; Nat. Med. 24, 50 (2018)

12. Cytotoxicity of propofol in human induced pluripotent stem cell-derived cardiomyocytes: K. Kido, et al.; J. Anesth. 32, 120 (2018), Application(s): Microplate reader

13. Development of novel amino-quinoline-5, 8-dione derivatives as NAD (P) H: quinone oxidoreductase 1 (NQO1) inhibitors with potent antiproliferative activities: Y. Ling, et al.; Eur. J. Med. Chem. 154, 199 (2018)

14. Inhibition of apoptosis using exosomes in Chinese hamster ovary cell culture: S. Han, et al.; Biotechnol. Bioeng. 115, 1331 (2018)

15. Light action spectrum on oxidative stress and mitochondrial damage in A2E-loaded retinal pigment epithelium cells: M. Marie, et al.; Cell Death Disc. 9, 287 (2018)

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