![]() |
产地 | American |
品牌 | AAT Bioquest |
货号 | CS23600 |
用途 | Research Grade |
包装规格 | 100 Tests |
纯度 | 98%% |
CAS编号 | |
是否进口 | 是 |
间隙连接是由连接蛋白构成的专门的细胞间膜通道,可选择性促进<1.5 kD的小分子通过细胞。它们受电压,生长因子,cAMP和类维生素A的紧密调节,并且受磷酸化调节。Cell Meter 荧光细胞间隙连接检测试剂盒为体外测定间隙连接功能提供了可靠而强大的检测方法。该方法是非侵入性的。对研究中的细胞群进行划分,以使其中一小部分载有亲脂性细胞质膜可渗透染料Calcein UltraGreen™AM,当细胞质酯酶吸收细胞时,该染料会水解,从而生成Calcein UltraGreen(一种高度荧光且保留良好的膜) -不可渗透的分子。另一部分是DiD,它是一种亲脂性膜染料,在掺入膜时会横向扩散,使整个细胞膜染成深红色荧光。将两个部分混合并在共培养条件下孵育。钙黄绿素UltraGreen通过间隙连接转移到DiD染色的细胞中。最终可以通过荧光成像或流式细胞术检测。
适用仪器
流式细胞仪 | |
激发: | 488 nm and 640 nm激光 |
发射: | 530/30 nm and 660/20 nm滤波片 |
通道: | FITC和APC通道 |
荧光显微镜 | |
激发: | FITC和Cy5滤波片组 |
发射: | FITC和Cy5滤波片组 |
推荐孔板: | 黑色透明 |
样品分析方案
概述
将钙黄绿素Ultragreen AM工作溶液添加到细胞中
将DiD工作溶液添加到细胞中
在37°C下孵育10-30分钟
用GAP连接检测缓冲液洗涤细胞
将细胞重悬于细胞培养基中并按1:1比例混合
使用带有530/30 nm和660/20 nm发射滤光片的流式细胞仪或带有FITC和Cy5滤光片组的荧光显微镜在不同时间检测荧光信号
储备溶液配制
操作步骤
图示
图1.通过流式细胞仪分析了GAP连接。根据方案,分别用钙黄绿素超绿AM和DiD对HeLa细胞进行染色。将细胞以1:1的比例充分混合,并用细胞培养基重新铺片。使用带有FITC和APC通道的NovoCyte流式细胞仪(ACEA Biosciences)检测反应。随着时间的推移,Q2-2(双阳性)增加。
参考文献
Cilostamide and forskolin maintain gap junction function of incubated dog follicles.
Authors: Thongkittidilok, Chommanart and Doriguzzi, Nicole and Nagashima, Jennifer and Brown, Megan and Chansaenroj, Ajjima and Songsasen, Nucharin
Journal: Theriogenology (2020): 222-228
Granulosa secreted factors improve the developmental competence of cumulus oocyte complexes from small antral follicles in sheep.
Authors: Rouhollahi Varnosfaderani, Shiva and Hajian, Mehdi and Jafarpour, Farnoosh and Ghazvini Zadegan, Faezeh and Nasr-Esfahani, Mohammad Hossein
Journal: PloS one (2020): e0229043
An Assay to Assess Gap Junction Communication in Cell Lines.
Authors: Warawdekar, Ujjwala M
Journal: Journal of biomolecular techniques : JBT (2019): 1-6
Cardiomyocyte-myofibroblast contact dynamism is modulated by connexin-43.
Authors: Schultz, Francisca and Swiatlowska, Pamela and Alvarez-Laviada, Anita and Sanchez-Alonso, Jose L and Song, Qianqian and de Vries, Antoine A F and Pijnappels, Dani?l A and Ongstad, Emily and Braga, Vania M M and Entcheva, Emilia and Gourdie, Robert G and Miragoli, Michele and Gorelik, Julia
Journal: FASEB journal : official publication of the Federation of American Societies for Experimental Biology (2019): 10453-10468
Cx43-Gap Junctions Accumulate at the Cytotoxic Immunological Synapse Enabling Cytotoxic T Lymphocyte Melanoma Cell Killing.
Authors: Hofmann, Francisca and Navarrete, Mariela and álvarez, Javiera and Guerrero, Israel and Gleisner, María Alejandra and Tittarelli, Andrés and Salazar-Onfray, Flavio
Journal: International journal of molecular sciences (2019)
G protein-coupled receptor 30 mediates meiosis resumption and gap junction communications downregulation in goat cumulus-oocyte complexes by 17β-estradiol.
Authors: Zhang, Hui and Wei, Qiang and Gao, Zhen and Ma, Chiyuan and Yang, Zhenshan and Zhao, Hui and Liu, Chen and Liu, Jie and Zhao, Xiaoe and Ma, Baohua
Journal: The Journal of steroid biochemistry and molecular biology (2019): 58-67
Perfluorooctanoic acid disrupts gap junction intercellular communication and induces reactive oxygen species formation and apoptosis in mouse ovaries.
Authors: López-Arellano, Patricia and López-Arellano, Keila and Luna, Jaquelinne and Flores, Diana and Jiménez-Salazar, Javier and Gavia, Graciela and Teteltitla, Mario and Rodríguez, Juan José and Domínguez, Alejandro and Casas, Eduardo and Bahena, Ivan and Betancourt, Miguel and González, Cristina and Ducolomb, Yvonne and Bonilla, Edmundo
Journal: Environmental toxicology (2019): 92-98
Gap junctions contribute to anchorage-independent clustering of breast cancer cells.
Authors: Gava, Fabien and Rigal, Lise and Mondesert, Odile and Pesce, Elise and Ducommun, Bernard and Lobjois, Valérie
Journal: BMC cancer (2018): 221
Heat stress impairs gap junction communication and cumulus function of bovine oocytes.
Authors: Campen, Kelly A and Abbott, Chelsea R and Rispoli, Louisa A and Payton, Rebecca R and Saxton, Arnold M and Edwards, J Lannett
Journal: The Journal of reproduction and development (2018): 385-392