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

關鍵詞：

DiSC3(5);DiOC5(3)；DiOC6(3)；ER-Tracker Blue-White DPX；Membrane depolarization assay膜去極化實驗；CAS：53213-94-8；

訂購信息：

產品描述

DiSC3(5) (3,3'-Dipropylthiadicarbocyanine Iodide)，一種具短（C3）烷基尾的羰花青染料，這種陽離子染料能用來檢測和測量由膜改性試劑引起的跨膜電位或結構變化。

產品特性

1. CAS NO：53213-94-8

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

3. 英文同義名：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. 中文同義名：3,3'-二丙基硫雜二羰花青碘化物；碘化-3,3ˊ-二丙基硫雜二羰花青；

5. 分子式：C25H27IN2S2

6. 分子量：546.53g/mol

7. 純度：≥98%（HPLC）

8.  Ex/Em：500/705 nm (in DMF)

9.  溶解性：溶于DMF

10.   化學結構式： 

保存與運輸方法

保存：室溫避光干燥保存，可置于-20℃長期干燥避光保存，2年有效。

運輸：室溫運輸。

應用示例：

文獻來源：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

實驗方法（膜去極化實驗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.

檢測原理（MK-DiSC3-(5)的膜去極化實驗）：DiSC3(5)作為一種膜電位敏感探針，聚集在磷脂雙分子層內，引起染料的自淬滅。當膜改性化合物（比如多肽）使膜去極化，電位喪失，DiSC3(5)釋放進入溶液引起熒光增強，熒光強弱與電位減少程度呈正比。 

結果展現（MK-DiSC3-(5)的膜去極化實驗）：

文獻來源：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

實驗方法（膜電位的顯微分析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)的工作濃度內維持1%DMSO濃度，對維持染料的溶解性和熒光穩定性很是關鍵?！?/p>

結果展現（MK-DiSC3-(5)的單細胞膜電位顯微分析）：

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。

相關產品：

——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。

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