• 主题
高级搜索  >
历史搜索 清空历史
首页> 外文期刊>Current Protocols in Cell Biology >The Sandwich Assay: A Methodfor Subcellular Visualizationof Paracellular Macromolecule Passagein Epithelial Sheets
【24h】

The Sandwich Assay: A Methodfor Subcellular Visualizationof Paracellular Macromolecule Passagein Epithelial Sheets

机译：三明治测定：上皮板中荚膜肺部大分子通道的亚细胞可视化方法

获取原文
获取原文并翻译 | 示例
           
页面导航
  • 摘要
  • 著录项
  • 相似文献
  • 相关主题

摘要

To date, the permeability of epithelia to larger solutes ( greater than 4Aindiameter) has been analyzed by ux measurements using various tracers thatcannot spatially resolve the permeation sites. This unit describes a method forlocalizing such sitesof passage in epithelial sheets with subcellular resolution.The method makes use of avidin as a basolateral capture probe in epithelialmonolayers or mucosae to unmask the passage of biotinylated and uorophore-labeled tracer molecules as they go throughthe junctional barrier. Once boundto avidin, the tracers are immobilized at the site of a barrier leak. The local-ization, the distribution, and the extent of passage are eventually evaluated byimaging. The assay detects single leaks and is hence able tospatially resolverarely occurring changes. It is also modular and exible to use with variousmacromolecular tracers, and its sensitivity is adjustable. If designed as a chaseexperiment, the method allows for analysis of temporal barrier openings. Ifperformed at low temperatures, this assay will block transcellular passage and,combined with global ux measurement, unambiguously determine paracellu-lar passage.
机译：迄今为止，通过使用各种示踪剂在空间地解析渗透位点，通过UX测量分析了上皮细胞对较大溶质的渗透率（大于4aindiameter）。该单元描述了一种用亚细胞分辨率在上皮片中通过的这种位点化的方法。该方法利用伊霉素作为上皮癌中的基础外侧捕获探针，以取代生物素化和鲁西氨光学标记的示踪剂分子的通过，因为它们通过连接屏障。一旦杀伤逃亡者，示踪剂就固定在屏障泄漏的部位。局部 - 局部，分布和通道的程度最终通过体析评估。测定检测单泄漏，因此能够在本旋地解决的变化。它也是可与各种摩托分子示踪剂一起使用的模块化且可用的，其灵敏度可调。如果设计为ChaseExperiment，该方法允许分析时间屏障开口。如果在低温下，该测定将阻断型横细胞通道，并结合全球UX测量，明确地确定帕拉库鲁 - LAR通道。

著录项

相似文献

  • 外文文献
  • 中文文献
  • 专利
获取原文

客服邮箱：kefu@zhangqiaokeyan.com

京公网安备：11010802029741号 ICP备案号：京ICP备15016152号-6 六维联合信息科技 (北京) 有限公司©版权所有

  • 客服微信

  • 服务号