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RNA结合蛋白的检测通过<em>体外</em在脂肪细胞培养> RNA下拉

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Biology

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JoVE Journal Biology

Detection of RNA-binding Proteins by In Vitro RNA Pull-down in Adipocyte Culture

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RNA结合蛋白的检测通过<em>体外</em在脂肪细胞培养> RNA下拉

DOI:

10.3791/54207-v

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10:34 min

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July 22, 2016

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Qianfan Bai*¹, Zhiqiang Bai*¹, Shaohai Xu, Lei Sun²

¹Cardiovascular and Metabolic Disorders Program,Duke-NUS Medical School, ²Institute of Molecular and Cell Biology, Singapore, ³Division of Bioengineering, School of Chemical & Biomedical Engineering,Nanyang Technological University

Chapters

00:05Title
00:35Preparation of Biotin-labeled RNA
02:18Preparation of RNA-conjugated Beads
04:16Preparation of Cellular Lysate from Primary Adipocytes
07:00Binding and Elution of RBP
08:45Results: RNA Pull-down Assay
10:05Conclusion

Summary

Automatic Translation

的RNA下拉协议这里优化用于检测RNA结合蛋白（限制性商业惯例）和非编码以及编码RNA之间的相互作用。从雄激素受体（AR）的RNA片段作为例子来演示如何从初级的棕色脂肪细胞的lystate检索其RBP。

Tags

RNA-binding Proteins RNA Pull-down Adipocyte Culture Antigen Receptor 3-Prime Untranslated Region (AR 3-Prime UTR)Biotinylated RNA In Vitro Transcription Streptavidin-coated Magnetic Beads RNA Structure Buffer RNA-protein Interactions RNA Regulatory Mechanisms Long Non-coding RNA

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