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

23.3K Views

10:34 min

July 22nd, 2016

July 22nd, 2016

23,304 Views

0:05

Title

0:35

Preparation of Biotin-labeled RNA

2:18

Preparation of RNA-conjugated Beads

4:16

Preparation of Cellular Lysate from Primary Adipocytes

7:00

Binding and Elution of RBP

8:45

Results: RNA Pull-down Assay

10:05

Conclusion

Transcript

The overall goal of this experiment is to retrieve RNA-binding proteins of an RNA transcript from brown adipose tissue. This method can help answer key questions in this field, such as, what are the regulatory mechanisms of longlife coding RNA, an

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An RNA pull-down protocol is optimized here for detection of interactions between RNA-binding proteins (RBPs) and noncoding as well as coding RNAs. An RNA fragment from androgen receptor (AR) was used as an example to demonstrate how to retrieve its RBP from lystate of primary brown adipocytes.

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