DiOC₂(3) 綠色膜電位熒光探針

產品關鍵詞：

DiOC2(3)；DiOC6(3)；壬基吖啶橙NAO；JC-10 線粒體膜電位探針；MitoTracker® Green FM 綠色線粒體熒光探針；羅丹明123 Rhodamine 123；CAS NO：905-96-4；

產品信息

【溫馨提示】：見我司懋康生物（maokangbio）提供的線粒體熒光探針產品專題。

產品描述

DiOC2(3)，英文全名3,3'-Diethyloxacarbocyanine Iodide，CAS NO. 905-96-4，是一種膜電位探針，可通過流式細胞儀檢測根據發射波長熒光信號比來分析細菌活力。DiOC2(3)常用來測定細菌膜電位，也能用于研究某些活細胞比如大鼠胚胎成纖維細胞、猴腎細胞、中國倉鼠肺成纖維細胞和小鼠3T3成纖維細胞。

DiOC2(3)用來檢測細菌膜電位的工作原理在于：DiOC2(3)在所有細菌細胞內發綠色熒光，由于更高的膜電位引起染料分子發生自聚作用，使得染料的熒光往紅色發射波長處遷移，紅色熒光強度增高，此時紅色/綠色熒光信號比高。質子離子載體比如CCCP，通過去除質子梯度來破壞線粒體膜電位，從而導致紅色熒光強度降低，此時紅色/綠色熒光信號比降低。DiOC2(3)的最大激發和發射波長約482nm和497nm。經DiOC2(3)標記的細胞用流式細胞儀分析（用488nm激發，分別用適合FITC（綠色）和Texas Red（紅色）的發射濾片來檢測）。

產品特性

1）   CAS NO.：905-96-4

2）   化學名：3-ethyl-2-[3-(3-ethyl-2(3H)-benzoxazolylidene)-1-propenyl]-benzoxazolium iodide

3）   同義名：3,3'-Diethyloxacarbocyanine Iodide 3,3′-二乙基氧雜羰花青碘; DiOC2(3) Iodide; DOC Iodide;

4）   分子式：C21H21IN2O2

5）   分子量：460.31

6）   純度：>95.0% (HPLC)

7）   Ex/Em：482/497nm（in methanol）

8）   外觀：紅色粉末或結晶

9）   溶解性：溶于DMSO，DMF，無水乙醇，微溶于水

10）化學結構式：

保存與運輸方法

保存：-20℃避光干燥保存，至少一年有效。

運輸：室溫運輸。

注意事項

1）   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。

2）   為了您的安全和健康，請穿實驗服并戴一次性手套操作。

使用方法

1.染色液制備

1）儲存液制備：用 DMSO或無水乙醇配置濃度1～5 mM的儲存液。例如，取10mg DiOC2(3)（Mw：460.31 g/mol）溶于4.35ml 無水DMSO中，充分溶解，即得到5mM的儲存液?！咀⒁狻课词褂玫膬Υ嬉悍盅b儲存在-20℃，避免反復凍融，且要避光保存。

2）工作液制備：用合適的緩沖液（如：無血清培養基，HBSS或PBS）稀釋儲存液，調整到合適的工作濃度?！咀⒁狻浚汗ぷ饕鹤罱K濃度需要根據不同細胞系和實驗體系來優化。

2.染色方法

對于細菌染色，最好取對數生長期的健康細菌，用無菌PBS或其他適當平衡鹽緩沖液稀釋到~1 x 106cells/ml。也可直接從細菌培養液中取適量的細菌不經清洗進行稀釋。細菌染色用DiOC2(3)的工作濃度可為30μM，室溫染色15~30min。

3.染色示例

Fig 1. Detection of membrane potential in mycobacteria. Red/green ratios were calculated using population mean fluorescence intensities for mycobacteria, incubated with 3 μM DiOC2 for 30 min in either the presence or absence of 25 μM CCCP.

相關產品

參考文獻

[1]Chen Z et al. Hypoionic Shock Facilitates Aminoglycoside Killing of Both Nutrient Shift- and Starvation-Induced Bacterial Persister Cells by Rapidly Enhancing Aminoglycoside Uptake. Front Microbiol. 2019 Sep 6;10:2028. PMID: 31551965

A flow cytometry-based assay was applied to measure the PMF by using the fluorescence probe 3,3′-Diethyloxacarbocyanine Iodide [DiOC2(3); purchased from MaoKang Biotechnology, Inc., Shanghai, China] according to the manufacturer’s instruction. Briefly, E. coli persisters, with or without CCCP pretreatment as described above, were diluted into PBS to a cell density of 106 cells/mL and incubated with DiOC2(3) (at a final concentration of 30 μM) at room temperature for 30 min. Cells were subjected to flow cytometric analysis on FACSymphonyTMA5 (BD Biosciences) with an excitation at 488 nm and emission at both red and green channels.

[2]Zhao Y et al. Rapid Freezing Enables Aminoglycosides To Eradicate Bacterial Persisters via Enhancing Mechanosensitive Channel MscL-Mediated Antibiotic Uptake. mBio. 2020 Feb 11;11(1). pii: e03239-19. doi: 10.1128/mBio.03239-19. PMID: 32047133

【3】A flow cytometry-based assay was applied to measure the proton motive force by using the fluorescence probe 3,3′-diethyloxacarbocyanine iodide [DiOC2(3)] (purchased from MaoKang Biotechnology, Inc., Shanghai, China) according to the manufacturer’s instructions. Briefly, E. coli exponential-phase cells, with or without CCCP pretreatment for 1 h, were diluted into PBS to a cell density of 106 cells/ml and incubated with DiOC2(3) (at a final concentration of 30?μM) at room temperature for 15?min. Cells were subjected to flow cytometric analysis on a FACSymphony A5 system (BD Biosciences) with excitation at 488?nm and emission in both the red (630-nm) channel and green (515-nm) channel. Cells treated by rapid freezing for 10?s with liquid nitrogen were also analyzed.

[4]Participation of the inositol 1,4,5-trisphosphate-gated calcium channel in the zona pellucida- and progesterone-induced acrosome reaction and calcium influx in human spermatozoa.

Hoechst 33258 and polyvinylpyrrolidone-40 (PVP-40) were obtained from Shanghai Maokang Biotechnology Co., Ltd. (Shanghai, China). The spermatozoa were washed and incubated with Hoechst 33258 (4 μg ml?1) for 10 min at a proportion of 9:1 to achieve a final concentration of 0.4 μg ml?1 and then centrifuged with 2% (w/v) PVP-40 in PBS. The supernatant was then removed and resuspended in PBS, smeared on clean slides, and fixed for 30 min in 95% (v/v) ethanol after air drying.

 — —Written/Edited by V. Shallan【版權歸MKBio懋康所有】

上海懋康生物科技有限公司是一家涉足于生命科學和生物技術領域研究的試劑、儀器和實驗室消耗品與實驗服務工作，主要從事細胞生物學、植物學、分子生物學、免疫學、生物化學、蛋白組學。生物制藥與診斷試劑研發生產等領域。 本公司秉承“以人為本，以誠為信、合同守信”的經營理念。堅持"品質保障"的原則為廣大客戶提供優質產品。