ER-Tracker Red (内质网红色荧光探针)(C1041)

ER-Tracker Red (内质网红色荧光探针)

产品编号: C1041

产品包装:20μl
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20μl

说明书下载

1258.00 10

价格: ¥ 1258.00

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产品编号 产品名称 产品包装 产品价格
C1041 ER-Tracker Red (内质网红色荧光探针) 20μl 1258.00元

ER-Tracker Red是一种具有细胞膜通透性的内质网(endoplasmic reticulum, ER)红色荧光探针,对内质网有高度选择性,可以用于活细胞内质网特异性荧光染色。
ER-Tracker Red为采用Molecular Probes公司的BODIPY TR进行了荧光标记的glibenclamide。Glibenclamide即glyburide,中文名为格列苯脲,是一种2型糖尿病治疗药物,可以结合主要定位在内质网上的包含ATP敏感的钾离子通道(KATP channel)的磺脲类(sulphonylurea)受体。因此荧光标记的glibenclamide就可以用作内质网特异性的荧光探针。ER-Tracker Red适用于活细胞内质网的荧光染色,但不适合用于固定细胞内质网的荧光染色。
ER-Tracker Red的分子式为C44H42BClF2N6O7S2,分子量为915.23,ER-Tracker Red的化学结构式参考图1。
ER-Tracker Red (内质网红色荧光探针)(C1041) 图1.ER-Tracker Red的化学结构式。 ER-Tracker Red对于细胞的毒性极低。而传统的DiOC6(3)对ER染色的同时也对细胞有一定的毒性。
ER-Tracker Red呈红色荧光,检测时的最大激发波长为587nm,最大发射波长为615nm。ER-Tracker Red的激发光谱和发射光谱参考图2。
ER-Tracker Red (内质网红色荧光探针)(C1041) 图2.ER-Tracker Red的激发光谱和发射光谱。 ER-Tracker Red按照后附的使用说明染色后,用甲醛等固定后染色效果可以被部分保留。ER-Tracker Red在活细胞中的染色效果参考图2。
ER-Tracker Red (内质网红色荧光探针)(C1041) 图3.ER-Tracker Red(内质网红色荧光探针)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细胞其线粒体呈现绿色荧光(图B),ER-Tracker Red染色的NRK-52E细胞其内质网呈现红色荧光(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。
提供了ER-Tracker Red 稀释液,使ER-Tracker Red的使用更加便捷。
按照1:1000的比例稀释,可以配制20ml ER-Tracker Red工作液;按照1:3000的比例稀释,可以配制60ml ER-TrackerRed工作液。
包装清单:

产品编号 产品名称 包装
C1041-1 ER-Tracker Red(1mM) 20μl
C1041-2 ER-Tracker Red稀释液 60ml
说明书 1份

保存条件:
-20℃保存,半年有效。ER-Tracker Red需-20℃避光保存。
注意事项:
ER-Tracker Red(1mM)在4℃、冰浴等较低温度情况下会凝固而粘在离心管管底、管壁或管盖内,可以20-25℃水浴温育片刻至全部融解后使用。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。
需自备盖玻片和载玻片(可以向碧云天订购)。
格列本脲的药理学活性可能会影响内质网的一些功能;某些特殊细胞中磺脲类受体的可变表达可能会导致非内质网特异性染色。
ER-Tracker Red适用于活细胞内质网的荧光染色,但不适合用于固定细胞内质网的荧光染色。如果经ER-Tracker Red染色后的细胞需要进行固定操作,使用4%多聚甲醛在37℃固定2分钟。
ER-Tracker Red染色后的细胞不能用Triton X-100通透,Triton X-100通透处理会导致ER-Tracker Red的荧光染色消失。
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.ER-Tracker Red工作液的配制:
a.取少量ER-Tracker Red按照1:1000的比例加入到ER-Tracker Red 稀释液中。例如取1μl ER-Tracker Red加入到1ml ER-Tracker Red 稀释液中。混匀后即为ER-Tracker Red工作液。
b.ER-Tracker Red工作液使用前需37℃预温育。
注:工作液中ER-Tracker Red的浓度可以根据实际情况进行适当调整,推荐的稀释比例调整范围为1:1000-1:3000。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ER-Tracker Red。
2.内质网的荧光标记:
a.去除细胞培养液,用适量的溶液如HBSS with Ca2+ & Mg2+ (Hanks’ Balanced Salt Solution with Ca2+ & Mg2+)洗涤生长在盖玻片上的细胞。注:HBSS with Ca2+ & Mg2+ (C0219)可以向碧云天订购;对于悬浮细胞的染色可以参考贴壁细胞的染色方法进行。
b.去除洗涤液,加入步骤1配制好的并37℃预温育的ER-Tracker Red染色工作液,与细胞37℃共孵育15-30分钟。
c.去除ER-Tracker Red染色工作液,用细胞培养液洗涤细胞1-2次。
d.随后通常用荧光显微镜或激光共聚焦显微镜进行观察。此时可观察到内质网呈明亮的强荧光染色。
e.如果经ER-Tracker Red染色后的细胞需要进行固定,可以使用4%甲醛37℃固定2分钟。固定后用适当的洗涤液洗涤2-3次,每次5分钟,随后可以进行复染或滴加适当的抗荧光淬灭封片液,最后封片观察。注意:ER-Tracker Red染色的细胞不能用Triton X-100通透,Triton X-100通透处理会导致ER-Tracker Red的荧光染色消失。

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ER-Tracker Red (内质网红色荧光探针)(C1041)

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