Mito-Tracker Green (线粒体绿色荧光探针)
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产品编号 C1048
数量
¥ 327.00
产品包装:
50μg
产品简介
使用说明
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Mito-Tracker Green,也称MitoTracker Green,是一种线粒体(mitochondria)特异性绿色荧光探针。Mito-Tracker Green可用于活细胞线粒体特异性荧光染色,也可用于固定细胞线粒体染色,但后者染色效果会略有下降。本产品染色后不能再进行固定。
Mito-Tracker Green为采用carbocyanine进行了荧光标记的一种Mito-Tracker,也称Benzoxazolium, 2-[3-[5,6-dichloro-1,3-bis[[4-(chloromethyl)phenyl]methyl]-1,3-dihydro-2H-benzimidazol-2-ylidene]-1-propenyl]-3-methyl-, chloride。分子式为C34H28Cl5N3O,分子量为671.88,CAS number为201860-17-5,可以用作线粒体特异性的荧光探针。和Mito-Tracker Deep Red 633、Mito-Tracker Deep Red FM、Rhodamine 123或JC-1相比,Mito-Tracker Green对于线粒体的染色不依赖于线粒体膜电位。
由于Mito-Tracker Green对于线粒体的染色不依赖于线粒体膜电位,因此可对固定后的细胞或组织进行染色,但荧光可能会有一定程度的下降或弥散。Mito-Tracker Green染色后固定后会导致荧光消失。Mito-Tracker Green对活细胞的染色效果参考图1。

图1.Mito-Tracker Green(线粒体绿色荧光探针)对于NRK-52E细胞(大鼠肾小管上皮细胞)的染色效果。Mito-Tracker Green染色的NRK-52E活细胞线粒体呈现绿色荧光(图B),Lyso-Tracker Red (C1046)染色的NRK-52E细胞的溶酶体呈现红色荧光(图C),绿色荧光、红色荧光及细胞核蓝色荧光的叠加(merge)效果见图D。其中细胞核使用Hoechst 33342 (C1027)染色。本图仅作参考,实际检测效果会因实验条件、检测仪器等的不同而存在差异。
Mito-Tracker Green最大激发光波长为490nm,最大发射波长为516nm,呈绿色荧光,Mito-Tracker Green的结构式以及激发光谱和发射光谱参考图2。

图2.Mito-Tracker Green的结构式(左)以及激发光谱和发射光谱(右)。

按最终工作浓度为20-200nM计算,可以配制约370-3700ml Mito-Tracker Green工作液。
包装清单:

产品编号 产品名称 包装
C1048 Mito-Tracker Green 50μg
说明书 1份

保存条件:
-20℃避光保存,半年有效。
注意事项:
活细胞染色后可以立即进行观察或检测,再经过固定后会导致荧光消失。如需染色后再固定,推荐Mito-Tracker Deep Red FM (线粒体深红荧光探针) (C1032)。Mito-Tracker Green可以对固定后的细胞或组织进行染色,但荧光染色效果可能会有一定程度的下降或弥散。如需进一步通透处理,推荐使用免疫染色通透液(Saponin) (P0095)。
对于微量的液体,每次使用前先离心数秒钟,使液体充分沉降到管底。
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。
需自备盖玻片载玻片(可以向碧云天订购)。
本产品仅限于专业人员的科学研究用,不得用于临床诊断或治疗,不得用于食品或药品,不得存放于普通住宅内。
为了您的安全和健康,请穿实验服并戴一次性手套操作。

使用说明:
1. Mito-Tracker Green储存液的配制:
用无水DMSO(anhydrous dimethylsulfoxide)配制Mito-Tracker Green至终浓度为1mM。配制后可以-20℃或更低温度避光保存。
2. Mito-Tracker Green工作液的配制:
a. 取少量1mM Mito-Tracker Green储存液按照1:5000-1:50000的比例加入到细胞培养液或适当的溶液(例如含钙镁离子的HBSS)中,使最终浓度为20-200nM。例如取1μl Mito-Tracker Green加入到50ml或5ml细胞培养液或适当的溶液(例如含钙镁离子的HBSS)中。混匀后即为Mito-TrackerGreen工作液。 HBSS with Ca2+ & Mg2+ (C0219)可以向碧云天订购。
b. Mito-Tracker Green工作液使用前需37℃预温育。
注:工作液中Mito-Tracker Green的浓度可以根据实际情况进行适当调整。为降低背景,在染色效果可以接受的范围内,建议尽量使用较低浓度的Mito-Tracker Green。
3. 线粒体的荧光标记:
a. 去除细胞培养液,加入步骤2配制好的并37℃预温育的Mito-Tracker Green染色工作液,与细胞37℃共孵育15-45分钟。
b. 去除Mito-Tracker Green染色工作液,加入37℃预温育的新鲜细胞培养液。
c. 随后通常用荧光显微镜或激光共聚焦显微镜进行观察。此时可观察到线粒体呈明亮的强荧光染色。如果染色效果欠佳,可以提高Mito-Tracker Green染色工作液中Mito-Tracker Green的浓度或在推荐的时间范围内适当延长染色时间。
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