| Chapter title |
Detecting RNA–Protein Interaction Using End-Labeled Biotinylated RNA Oligonucleotides and Immunoblotting
|
|---|---|
| Chapter number | 4 |
| Book title |
RNA-Protein Complexes and Interactions
|
| Published in |
Methods in molecular biology, January 2016
|
| DOI | 10.1007/978-1-4939-3591-8_4 |
| Pubmed ID | |
| Book ISBNs |
978-1-4939-3589-5, 978-1-4939-3591-8
|
| Authors |
Xuexiu Zheng, Sunghee Cho, Heegyum Moon, Tiing Jen Loh, Ha Na Jang, Haihong Shen, Zheng, Xuexiu, Cho, Sunghee, Moon, Heegyum, Loh, Tiing Jen, Jang, Ha Na, Shen, Haihong |
| Abstract |
RNA-protein interaction can be detected by RNA pull-down and immunoblotting methods. Here, we describe a method to detect RNA-protein interaction using RNA pull down and to identify the proteins that are pulled-down by the RNA using immunoblotting. In this protocol, RNAs with specific sequences are biotinylated and immobilized onto Streptavidin beads, which are then used to pull down interacting proteins from cellular extracts. The presence of a specific protein is subsequently verified by SDS- polyacrylamide gel electrophoresis and immunoblotting with antibodies. Interactions between the SMN RNA and the PSF protein and between the caspase-2 RNA and the SRSF3 protein (SRp20) in nuclear extract prepared from HeLa cells are illustrated as examples. |
Mendeley readers
Geographical breakdown
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|---|---|---|
| Unknown | 49 | 100% |
Demographic breakdown
| Readers by professional status | Count | As % |
|---|---|---|
| Student > Ph. D. Student | 11 | 22% |
| Student > Bachelor | 7 | 14% |
| Student > Master | 7 | 14% |
| Researcher | 4 | 8% |
| Professor | 3 | 6% |
| Other | 3 | 6% |
| Unknown | 14 | 29% |
| Readers by discipline | Count | As % |
|---|---|---|
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| Chemistry | 3 | 6% |
| Immunology and Microbiology | 3 | 6% |
| Pharmacology, Toxicology and Pharmaceutical Science | 1 | 2% |
| Other | 3 | 6% |
| Unknown | 14 | 29% |