用于量化连接蛋白在固定上皮细胞中的水平和垂直分布的协议
Madeline Lovejoy1, Claudio A Mucci1, Agustín Rabino1
1Department of Molecular, Cellular, and Developmental Biology, University of Toledo, Toledo, OH 43606, USA.
STAR protocols
|September 12, 2025
概括
这项研究引入了一种可复制的协议,用于量化上皮细胞中的结节蛋白. 它详细介绍了使用斐济/ImageJ和RStudio的样品制备,免疫光和图像分析方法.
科学领域:
- 细胞生物学 细胞生物学
- 生物物理学的生物物理.
背景情况:
- 两极化的上皮细胞形成密集的单层,对组织完整性至关重要.
- 细胞-细胞结合,包括附着结合 (AJs) 和紧密结合 (TJs),调解细胞粘附和屏障功能.
- 了解结节蛋白的分布是破译上皮质细胞行为的关键.
研究的目的:
- 为量化连接蛋白的水平和垂直分布提出一个详细的协议.
- 为了能够在不同的实验条件下对结点蛋白分布进行比较分析.
- 为没有图像分析软件经验的研究人员提供可访问的方法.
主要方法:
- 合表皮细胞单层的样本制备和固定.
- 针对特定的结点蛋白质进行免疫光染色.
- 使用显微镜获取图像,并使用斐济/ImageJ和RStudio.com进行定量分析.
主要成果:
- 一个用于可视化和量化结点蛋白质分布的综合性协议.
- 展示如何在水平和垂直平面上分析蛋白质定位.
- 适用于各种实验条件和细胞类型的方法.
结论:
- 本协议提供了一种标准化和可访问的方法来研究结点蛋白质动力学.
- 这种方法有助于在实验中对表皮细胞结合点进行定量比较.
- 能够更深入地了解细胞-细胞结合的结构组织和调节.
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