Cell Navigator™TMR神经酰胺高尔基体染色试剂盒*红色荧光*

货号22752存储条件 在零下15度以下保存, 避免光照
规格100 Tests价格3648
Ex (nm)544Em (nm)570
分子量溶剂
产品详细介绍


简要概述

高尔基体是大多数真核细胞胞质内囊泡和折叠膜的复合体,参与分泌和细胞内运输。它修饰内质网(ER)中内置的蛋白质和脂质,并准备将其运输到细胞外。它还在脂质在整个细胞中的运输和溶酶体的形成中起着重要作用。Cell Navigator™TMR神经酰胺高尔基染色试剂盒提供了一种简便,快速的方法,可以在活细胞中以红色荧光染色高尔基体。高尔基体通过形成相应的荧光代谢产物而被染色。Cell Navigator™TMR神经酰胺高尔基染色试剂盒提供了一种优化的测定方法,可用于通过荧光显微镜检查高尔基体的形态。

点击查看光谱

 

适用仪器


荧光显微镜  
激发: Cy3/TRITC滤波片组
发射: Cy3/TRITC滤波片组
推荐孔板: 黑色透明
 


产品说明书

样品分析方案

概述

根据需要处理细胞

加入GGR169-神经酰胺工作溶液并在室温或37°C下孵育15至30分钟

添加染色缓冲液

使用Cy3滤波片组在显微镜下观察 

 

储备溶液配制

GGR169-神经酰胺原液(100X)
向GGR-神经酰胺(组分A)中加入100 µL DMSO(组分C)制成GGR169-神经酰胺原液(100X)。
注意:将未使用的GGR-神经酰胺原液分装在-20°C下,以单次使用,避免冻融循环。
 
工作溶液配制
GGR169-神经酰胺工作液
将10 µL GGR169-神经酰胺原液(100X)添加到990 µL染色缓冲液(组分B)中,制成GGR169-神经酰胺工作液。
可选:向1 mL GGR169-神经酰胺工作溶液中加入10 µL Hoechst 33342(组分D)以进行核染色。在带有DAPI滤光片组的荧光显微镜下观察。

 

操作步骤

以下指南应用于指导方针,并可根据要求进行修改。
  1. 接种并根据需要处理细胞。
  2. 将100 µL GGR169-神经酰胺工作溶液直接添加到细胞培养基中。
  3. 在室温或37°C下孵育15至30分钟。
  4. 除去GGR169-神经酰胺工作溶液,并用DPBS或您选择的缓冲液清洗一次。
  5. 加入100 µL /孔的染色缓冲液(组分B)。
  6. 在装有Cy3滤光片的荧光显微镜下观察。 

 

图示

图1.HeLa细胞中GGR169神经酰胺高尔基染色的荧光图像。在37°C下,用100μL具有Hoechst 33342的GGR169-神经酰胺工作溶液对细胞染色20分钟。

 

参考文献

Chlamydia trachomatis-infected human cells convert ceramide to sphingomyelin without sphingomyelin synthases 1 and 2.
Authors: Tachida, Yuriko and Kumagai, Keigo and Sakai, Shota and Ando, Shuji and Yamaji, Toshiyuki and Hanada, Kentaro
Journal: FEBS letters (2020): 519-529

The role of ceramide in regulating endoplasmic reticulum function.
Authors: Zelnik, Iris D and Ventura, Ana E and Kim, Jiyoon L and Silva, Liana C and Futerman, Anthony H
Journal: Biochimica et biophysica acta. Molecular and cell biology of lipids (2020): 158489

Neuronal ceroid lipofuscinosis related ER membrane protein CLN8 regulates PP2A activity and ceramide levels.
Authors: Adhikari, Babita and De Silva, Bhagya and Molina, Joshua A and Allen, Ashton and Peck, Sun H and Lee, Stella Y
Journal: Biochimica et biophysica acta. Molecular basis of disease (2019): 322-328

Structure, functions and regulation of CERT, a lipid-transfer protein for the delivery of ceramide at the ER-Golgi membrane contact sites.
Authors: Kumagai, Keigo and Hanada, Kentaro
Journal: FEBS letters (2019): 2366-2377

Activation of neutral sphingomyelinase 2 by starvation induces cell-protective autophagy via an increase in Golgi-localized ceramide.
Authors: Back, Moon Jung and Ha, Hae Chan and Fu, Zhicheng and Choi, Jong Min and Piao, Yongwei and Won, Jong Hoon and Jang, Ji Min and Shin, In Chul and Kim, Dae Kyong
Journal: Cell death & disease (2018): 670

Antidepressants act by inducing autophagy controlled by sphingomyelin-ceramide.
Authors: Gulbins, Anne and Schumacher, Fabian and Becker, Katrin Anne and Wilker, Barbara and Soddemann, Matthias and Boldrin, Francesco and Müller, Christian P and Edwards, Michael J and Goodman, Michael and Caldwell, Charles C and Kleuser, Burkhard and Kornhuber, Johannes and Szabo, Ildiko and Gulbins, Erich
Journal: Molecular psychiatry (2018): 2324-2346

Both the N- and C- terminal regions of the Chlamydial inclusion protein D (IncD) are required for interaction with the pleckstrin homology domain of the ceramide transport protein CERT.
Authors: Kumagai, Keigo and Elwell, Cherilyn A and Ando, Shuji and Engel, Joanne N and Hanada, Kentaro
Journal: Biochemical and biophysical research communications (2018): 1070-1076

Ceramide Transporter CERT Is Involved in Muscle Insulin Signaling Defects Under Lipotoxic Conditions.
Authors: Bandet, Cécile L and Mahfouz, Rana and Véret, Julien and Sotiropoulos, Athanassia and Poirier, Maxime and Giussani, Paola and Campana, Mélanie and Philippe, Erwann and Blachnio-Zabielska, Agnieszka and Ballaire, Raphaëlle and Le Liepvre, Xavier and Bourron, Olivier and Berkeš, Dušan and Górski, Jan and Ferré, Pascal and Le Stunff, Hervé and Foufelle, Fabienne and Hajduch, Eric
Journal: Diabetes (2018): 1258-1271

Ceramide-transfer protein-mediated ceramide transfer is a structurally tunable flow-inducing mechanism with structural feed-forward loops.
Authors: Giordano, Giulia
Journal: Royal Society open science (2018): 180494

Phosphoinositide binding by the PH domain in ceramide transfer protein (CERT) is inhibited by hyperphosphorylation of an adjacent serine-repeat motif.
Authors: Sugiki, Toshihiko and Egawa, Daichi and Kumagai, Keigo and Kojima, Chojiro and Fujiwara, Toshimichi and Takeuchi, Koh and Shimada, Ichio and Hanada, Kentaro and Takahashi, Hideo
Journal: The Journal of biological chemistry (2018): 11206-11217



说明书
Cell Navigator™TMR神经酰胺高尔基体染色试剂盒*红色荧光*.pdf