尼罗红
中文名称 | 尼罗红 |
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中文同义词 | 尼罗红, 用于荧光分析;尼罗红5H-BENZO[A]PHENOXAZIN-5-ONE,9-(DIETHYLAMINO)-;Nile Red, 用于荧光分析,≥98.0% (HPLC);尼罗红, 用于荧光分析,≥98.0% (HPLC);Nile Red, 用于荧光分析,≥95.0% (HPLC);尼罗红, 用于荧光分析,≥95.0% (HPLC);尼罗红/尼罗丝/NILE RED;尼罗红 500MG |
英文名称 | Nile Red |
英文同义词 | NILE BLUE A OXAZONE;NILE RED;9-(DIETHYLAMINO)-5H-BENZO[A]PHENOXAZIN-5-ONE;9-(diethylamino)-5h-benzo[a]phenoxazin-5-on;nile;9-(diethylamino)benzo[a]phenoxazin-5(5H)-one;FLUORESCENCE QY-STANDARD E CERTIFIED;Fluorescent Spectral Standard E, certified |
CAS号 | 7385-67-3 |
分子式 | C20H18N2O2 |
分子量 | 318.37 |
EINECS号 | 230-966-0 |
相关类别 | 化工原料;生化试剂-色素类;其他生化试剂;色素类;生化试剂;Electroluminescence;Functional Materials;marker;荧光探针,标记,颗粒和染色;通用生化试剂-生物染色剂与化学指示剂;有机化工原料 |
Mol文件 | 7385-67-3.mol |
结构式 | ![]() |
尼罗红 性质
熔点 | 203-205 °C(lit.) |
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沸点 | 457.53°C (rough estimate) |
密度 | 1.1871 (rough estimate) |
折射率 | 1.5700 (estimate) |
储存条件 | 2-8°C |
溶解度 | 在甲醇中可溶1mg/mL |
形态 | 粉末 |
酸度系数(pKa) | 4.08±0.20(Predicted) |
颜色 | 深红色至栗色 |
外观 | Green to Dark Green Solid |
最大波长(λmax) | 553 nm |
BRN | 279110 |
稳定性 | 稳定的。与强氧化剂不相容。 |
InChI | InChI=1S/C20H18N2O2/c1-3-22(4-2)13-9-10-16-18(11-13)24-19-12-17(23)14-7-5-6-8-15(14)20(19)21-16/h5-12H,3-4H2,1-2H3 |
InChIKey | VOFUROIFQGPCGE-UHFFFAOYSA-N |
SMILES | C12=CC=CC=C1C(=O)C=C1C2=NC2=C(C=C(N(CC)CC)C=C2)O1 |
CAS 数据库 | 7385-67-3(CAS DataBase Reference) |
EPA化学物质信息 | 5H-Benzo[a]phenoxazin-5-one, 9-(diethylamino)- (7385-67-3) |
2)染色步骤
① 尼罗红染色液配制:a.母液准备:取适量的尼罗红(Mw:318.37g/mol)用无水DMSO充分溶解制备1mM储存液,按照单次用量分装冻存,避免反复冻融,避光保存。b.工作液准备:用HHBS或生理缓冲液(pH 7.0)将母液按1:1000稀释为1×尼罗红工作液,漩涡混匀。
② 用检测化合物处理细胞一段时间;
③ 离心并调整细胞浓度为1-5×10^5cell/管;
④ 用500µL尼罗红工作液重悬细胞;
⑤ 室温或37ºC避光孵育5-10min;
⑥ 吸掉染色工作液,用HHBS或适当缓冲液清洗细胞;
⑦ 用500µL预热的HHBS或培养基重悬细胞,使细胞密度为1-5×105cell/管;
⑧ 荧光显微镜或流式细胞仪检测荧光信号,Ex/Em=552/636nm。
【注意】:① 对于贴壁细胞,可用HHBS或适当缓冲液清洗细胞,然后直接加入尼罗红工作液孵育;② 细胞也可预固定,然后用尼罗红工作液染色。Nile Red (Nile Blue A oxazone, Phenoxazone 9)是一种十分有效的荧光染料,可在疏水环境下检测细胞内脂质滴 intracellular lipid droplets。Nile Red 可用于细胞内脂质,蛋白质的疏水域和溶酶体磷脂内含体的染色。
Target | Value |
lipid droplet
() |
Nile red-stained, lipid droplet-filled macrophages exhibit greater fluorescence intensity than does Nile red-stained control macrophages, and the two cell populations could be differentiated and analyzed by flow cytofluorometry. Better selectivity for cytoplasmic lipid droplets is obtained when the cells are viewed for yellow-gold fluorescence (excitation, 450-500 nm; emission, greater than 528 nm) rather than red fluorescence (excitation, 515-560 nm; emission, greater than 590 nm).
Nile red is strongly fluorescent, but only in the presence of a hydrophobic environment. Nile red is very soluble in the lipids it is intended to show, and it does not interact with any tissue constituent except by solution.
Spectral and physicochemical properties of the lipophilic dye Nile red induce a yellow-gold-spectral shift in its excitation-emission peak, allowing it to fluoresce in the green emission spectrum only when in a lipid-rich environment, but not in more polar environments.
When Nile red-stained Caenorhabditis elegans is viewed for green fluorescence, discrete lipid bodies can be observed throughout the intestine and other tissues either in clusters or evenly dispersed, depending on the animal's genotype or experimental treatment.
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