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

產(chǎn)品簡介：

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(jié)構(gòu)變化。

產(chǎn)品特性：

1） CAS NO：53213-94-8

2） 化學(xué)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

保存條件: 

室溫避光干燥保存，可置于-20℃長期干燥避光保存，2年有效。

產(chǎn)品使用：

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）：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（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(nèi)，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(jié)果展現(xiàn)（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）：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(nèi)維持1%DMSO濃度，對維持染料的溶解性和熒光穩(wěn)定性很是關(guān)鍵?！?/span>

結(jié)果展現(xiàn)（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。

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