DiSC3(5) (3,3'-Dipropylthiadicarbocyanine Iodide)

关键词：

DiSC3(5);DiOC5(3)；DiOC6(3)；ER-Tracker Blue-White DPX；Membrane depolarization assay膜去极化实验；CAS：53213-94-8；

订购信息：

产品描述

DiSC3(5) (3,3'-Dipropylthiadicarbocyanine Iodide)，一种具短（C3）烷基尾的羰花青染料，这种阳离子染料能用来检测和测量由膜改性试剂引起的跨膜电位或结构变化。

产品特性

1. CAS NO：53213-94-8

2. 化学名：Benzothiazolium, 3-propyl-2-(5-(3-propyl)-2(3H)-benzothiazolidene-1,3-pentadienyl), iodide

3. 英文同义名：diS-C3-(5); 3,3'-Di-n-propylthiadicarbocyanine iodide; 3,3-Dipropylthiadicarbocyanine iodide; 5-(3-Propylbenzothiazol-2-ylidene)-1-(3-propylbenzothiazolium-2-yl)-1,3-pentadiene Iodide; 3-Propyl-2-[5-[3-propyl-2(3H)-benzothiazolylidene]-1,3-pentadienyl]benzothiazolium iodide;

4. 中文同义名：3,3'-二丙基硫杂二羰花青碘化物；碘化-3,3ˊ-二丙基硫杂二羰花青；

5. 分子式：C25H27IN2S2

6. 分子量：546.53g/mol

7. 纯度：≥98%（HPLC）

8.  Ex/Em：500/705 nm (in DMF)

9.  溶解性：溶于DMF

10.   化学结构式： 

保存与运输方法

保存：室温避光干燥保存，可置于-20℃长期干燥避光保存，2年有效。

运输：室温运输。

应用示例：

文献来源：Juba ML et al. Helical cationic antimicrobial peptide length and its impact on membrane disruption. Biochim Biophys Acta. 2015 May;1848(5):1081-91. PMID: 25660753

实验方法（膜去极化实验MK）：Frozen aliquots ofenumerated bacteria (E. coli or B. cereus)were thawed on ice and washed 3 times with buffer (5 mM HEPES with 20 mM glucose, pH 7.4). Following washing, the pelleted bacteria were re-suspended in HEPES buffer (5 mM HEPES, pH 7.4, 20 mM glucose) containing either, 10 or 100 mM KCl. A 96-well plate was prepared where wells are charged with360 μL of bacterial suspension (2 × 107 CFU/mL) and 4.19 μL of diSC3-(5) (200 nM) for a total volume of 364.19 μL.The bacteria were incubated at room temperature and fluorescence was monitored (622 nmex/670 nmem) until diSC3-(5) maximal uptake was obtained. Maximal diSC3-(5) uptake is indicated by a baseline in fluorescence due to self-quenching as the dye concentrates in the cell membrane. Peptide (NA-CATH, L- or D-ATRA-1A) was added at varied concentrations (200–2 μg/mL) in 20 μL aliquots and the fluorescence increase due to induced depolarization of the cytoplasmic membrane was recorded. A negative control of bacteria and diSC3-(5) was used as a background. As a positive control, complete collapse of the membrane potential was attained with valinomycin (200–2 μg/mL), a potassium ionophore.

检测原理（MK-DiSC3-(5)的膜去极化实验）：DiSC3(5)作为一种膜电位敏感探针，聚集在磷脂双分子层内，引起染料的自淬灭。当膜改性化合物（比如多肽）使膜去极化，电位丧失，DiSC3(5)释放进入溶液引起荧光增强，荧光强弱与电位减少程度呈正比。 

结果展现（MK-DiSC3-(5)的膜去极化实验）：

文献来源：Te Winkel JDet al.Analysis of Antimicrobial-Triggered Membrane Depolarization Using Voltage Sensitive Dyes. Front Cell Dev Biol. 2016 Apr 13;4:29. PMID: 27148531

实验方法（膜电位的显微分析MK）：Early-mid logarithmic growth phase cell suspensions were incubated with 2 μM DiSC3(5)directly in the growth medium. The incubation was carried out under shaking for 5 min, immediately followed by microscopy.A final concentration of 0.5–1% DiSC3(5)-solvent dimethyl sulfoxide (DMSO) was found to be crucial in order to maintain appropriate solubility;lower solvent concentrations resulted in a strongly reduced cellular fluorescence. The incubation was carried out at growth temperature, and under vigorous shaking in order to maintain good energization of the cells. This step was routinely carried out with 2 ml round bottom Eppendorf tubes containing 200 μl cell suspensionin a thermomixer. To provide sufficient aeration, the lids of the tubes were perforated. When dissipation of membrane potential was tested, a compound of interest was added in parallel to DiSC3(5).Addition of 5 μM gramicidin (a mixture of gramicidin A, B, C, and D)was routinely used as a positive control. The imaging of DiSC3(5)-stained cells was carried out using commonly availableCy5-filter sets.【重要信息：文章多次提及到DiSC3(5)的工作浓度内维持1%DMSO浓度，对维持染料的溶解性和荧光稳定性很是关键。】

结果展现（MK-DiSC3-(5)的单细胞膜电位显微分析）：

注意事项

1）荧光染料均存在淬灭问题，请尽量注意避光，以减缓荧光淬灭。

2） 为了您的安全和健康，请穿实验服并戴一次性手套操作。

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——Written/Edited by V. Shallan【版权归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学和生物技术领域研究的试剂、仪器和实验室消耗品与实验服务工作，主要从事细胞生物学、植物学、分子生物学、免疫学、生物化学、蛋白组学。生物制药与诊断试剂研发生产等领域。 本公司秉承“以人为本，以诚为信、合同守信”的经营理念。坚持"品质保障"的原则为广大客户提供优质产品。

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