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利用邻近结扎测定法在人类胰腺β细胞中原位内源性米托巴结物的可视化

JoVE Journal
Medicina

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JoVE Journal Medicina

Visualization of Endogenous Mitophagy Complexes In Situ in Human Pancreatic Beta Cells Utilizing Proximity Ligation Assay

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利用邻近结扎测定法在人类胰腺β细胞中原位内源性米托巴结物的可视化

DOI:

10.3791/59398-v

•

08:40 min

•

May 02, 2019

•

Gemma Pearson, Scott A. Soleimanpour²

¹Department of Internal Medicine, Division of Metabolism, Endocrinology and Diabetes,University of Michigan, Ann Arbor, ²VA Ann Arbor Healthcare System

Capítulos

00:04Título
00:32Human Islet Sample Single Cell Dissociation
02:14Single Cell Adherence and Fixation and Immunohistochemistry
03:24Proximity Ligation Assay (PLA)
04:38Imaging
06:00PLA Interaction Quantification
07:18Results: Representative Beta Cell Mitophagy Analysis
08:08Conclusion

Summary

Tadução automática

该协议概述了一种定量分析从原人类小岛样本的β细胞中专门形成米托巴细胞蛋白复合体的方法。因此,该技术允许从有限的生物材料分析米托巴,这在珍贵的人类胰腺β细胞样本中至关重要。

Tags

Mitophagy Proximity Ligation Assay Human Pancreatic Beta Cells Endogenous Mitophagy Complexes Immunohistochemistry Ubiquitin-specific Peptidase 8 Neuregulin Receptor Degradation Protein-1 Beta Cell Marker

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