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缺氧条件下培养细胞内源性核蛋白的共免疫沉淀测定

JoVE Journal
Biologia

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JoVE Journal Biologia

Co-immunoprecipitation Assay Using Endogenous Nuclear Proteins from Cells Cultured Under Hypoxic Conditions

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缺氧条件下培养细胞内源性核蛋白的共免疫沉淀测定

DOI:

10.3791/57836-v

•

09:17 min

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August 02, 2018

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Xiaofeng Zheng², Calvin Qing Wei Ho, Xiaowei Zheng, Kian Leong Lee, Katarina Gradin, Teresa S. Pereira, Per-Olof Berggren^2,3, Yusuf Ali²

¹Lee Kong Chian School of Medicine,Nanyang Technological University, ²Singapore Eye Research Institute (SERI),Singapore General Hospital, ³The Rolf Luft Research Center for Diabetes and Endocrinology, Karolinska Institutet,Karolinska University Hospital, ⁴Cancer and Stem Cell Biology Program,Duke-NUS Medical School, ⁵Department of Cell and Molecular Biology,Karolinska Institutet

Capítulos

00:04Título
00:33Induction of Hypoxia in HEK293A Cells
01:28Harvesting Cells Cultured Under Hypoxic Conditions
03:30Preparing Nuclear Extracts
05:52Immunoprecipitation of Proteins
07:55Results: HIF-1α Interacts with ARNT at Hypoxia
08:56Conclusion

Summary

Tadução automática

在这里, 我们描述了一个共同免疫沉淀协议, 研究在缺氧条件下内源核蛋白的蛋白质相互作用。该方法适用于低氧转录因子与转录联合调节剂相互作用的证明。

Tags

Keywords: Co-immunoprecipitation Endogenous Nuclear Proteins Hypoxic Conditions Transcription Factors Transcriptional Coregulators Human Embryonic Kidney 293A Cells DPBS Hypoxia Subchamber Normoxia Cell Culture Cell Harvesting Nuclear Fractions

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