Naph-EA-mal
(Synonyms: Thiol-green 1) 目录号 : GC61556Naph-EA-mal(Thiol-green1)是一种快速检测和超快turn-on型硫醇(thiol)荧光探针,可用于蛋白标记和生物成像。Naph-EA-mal(Thiol-green1)可用于检测活细胞中的硫醇,标记蛋白质硫醇,定量细胞裂解液中总硫醇的浓度,并确定固定细胞中可逆的蛋白质硫醇氧化。Ex:488nM;Em:540nM。
Cas No.:210292-65-2
Sample solution is provided at 25 µL, 10mM.
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Naph-EA-mal (Thiol-green 1) is a rapid detect and ultrafast turn-on thiol fluorescence probe for protein labeling and bioimaging. Naph-EA-mal (Thiol-green 1) can be used to detect thiols in living cells, label the protein thiols, quantify the concentration of total thiols in cell lysate, and determine the reversible protein thiols oxidation in fixed cells[1]. Ex: 488 nM; Em: 540 nM.
Imaging thiols in living cells[1]Hep G2 cells incubated with Naph-EA-mal (Thiol-green 1) (1 µM;5 mins), and strong green fluorescence can be observed. When cells are preincubated the cells to block the intracellular thiols with NEM (a thiol block reagent; 100 µM), a weaker fluorescence is observed in cells[1].Monitoring thiol-disulfide levels in fixed cellsDue to the majority of cellular thiols are present in the reduced form, Naph-EA-mal (Thiol-green 1) (10 µM) induces strong fluorescence in Hep G2 cells. After diamide treatment, fluorescent signal sharp dimed is observed[1].The Hep G2 cells, oxidized by diamide (200 µM; 30 min), are fixed by 70% EtOH and the remaining free thiols are blocked by NEM (100 µM). then The cells are further treated with TCEP (5 mM) followed by addition of the Naph-EA-mal (Thiol-green 1) (10 µM). The green fluorescent signal is detected again[1]
[1]. Jinyu Sun, et al. An ultrafast turn-on thiol probe for protein labeling and bioimaging. Analyst
Cas No. | 210292-65-2 | SDF | |
别名 | Thiol-green 1 | ||
Canonical SMILES | O=C1N(CCCC)C(C2=CC=C(NCCN3C(C=CC3=O)=O)C4=CC=CC1=C24)=O | ||
分子式 | C22H21N3O4 | 分子量 | 391.42 |
溶解度 | DMSO : 33.33 mg/mL (85.15 mM; Need ultrasonic) | 储存条件 | Store at -20°C |
General tips | 请根据产品在不同溶剂中的溶解度选择合适的溶剂配制储备液;一旦配成溶液,请分装保存,避免反复冻融造成的产品失效。 储备液的保存方式和期限:-80°C 储存时,请在 6 个月内使用,-20°C 储存时,请在 1 个月内使用。 为了提高溶解度,请将管子加热至37℃,然后在超声波浴中震荡一段时间。 |
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制备储备液 | |||
1 mg | 5 mg | 10 mg | |
1 mM | 2.5548 mL | 12.774 mL | 25.548 mL |
5 mM | 0.511 mL | 2.5548 mL | 5.1096 mL |
10 mM | 0.2555 mL | 1.2774 mL | 2.5548 mL |
第一步:请输入基本实验信息(考虑到实验过程中的损耗,建议多配一只动物的药量) | ||||||||||
给药剂量 | mg/kg | 动物平均体重 | g | 每只动物给药体积 | ul | 动物数量 | 只 | |||
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% DMSO % % Tween 80 % saline | ||||||||||
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工作液浓度: mg/ml;
DMSO母液配制方法: mg 药物溶于 μL DMSO溶液(母液浓度 mg/mL,
体内配方配制方法:取 μL DMSO母液,加入 μL PEG300,混匀澄清后加入μL Tween 80,混匀澄清后加入 μL saline,混匀澄清。
1. 首先保证母液是澄清的;
2.
一定要按照顺序依次将溶剂加入,进行下一步操作之前必须保证上一步操作得到的是澄清的溶液,可采用涡旋、超声或水浴加热等物理方法助溶。
3. 以上所有助溶剂都可在 GlpBio 网站选购。
An ultrafast turn-on thiol probe for protein labeling and bioimaging
Analyst 2016 Mar 21;141(6):2009-15.PMID:26886182DOI:10.1039/c5an02347e
A novel turn-on type of ultrafast biothiol fluorescent probe, Naph-EA-mal, was designed, synthesized and evaluated. The probe contains a naphthalimide moiety as a fluorophore, a maleimide unit as a thiol acceptor, and 1,2-ethylenediamine as a linker. Naph-EA-mal displays high selectivity and a fast response toward thiols in aqueous solution. The reaction mechanism of the probe with thiols was confirmed by 1H NMR and HRMS. Test strips were fabricated and a sharp color change was observed by the naked-eye. Furthermore, Naph-EA-mal was successfully applied to label protein thiols, image thiols in living cells, quantify thiol content in cells lysate, and determine the reversible protein thiols oxidation in fixed cells.