| Chapter title |
Isolation and Characterization of Endogenous RNPs from Brain Tissues
|
|---|---|
| Chapter number | 28 |
| Book title |
RNA Detection
|
| Published in |
Methods in molecular biology, January 2018
|
| DOI | 10.1007/978-1-4939-7213-5_28 |
| Pubmed ID | |
| Book ISBNs |
978-1-4939-7212-8, 978-1-4939-7213-5
|
| Authors |
Rico Schieweck, Foong yee Ang, Renate Fritzsche, Michael A. Kiebler, R Schieweck, FY Ang, R Fritzsche, MA Kiebler |
| Abstract |
Identification of physiological target RNAs and protein interactors bound to RNA-binding proteins is a key prerequisite to understand the underlying mechanisms of posttranscriptional expression control and RNA granule assembly. Here, we describe a multistep biochemical approach to isolate endogenous ribonucleoprotein particles from brain tissues by exploiting differential centrifugation and gradient fractionation followed by immunoprecipitation with monospecific, affinity-purified antibodies directed against selected RNA-binding proteins. This protocol results in highly enriched endogenous ribonucleoprotein particles that then can be analyzed by mass spectrometry (for proteins composition) and microarray or RNA sequencing technologies (for target mRNAs). |
X Demographics
Geographical breakdown
| Country | Count | As % |
|---|---|---|
| United Kingdom | 1 | 100% |
Demographic breakdown
| Type | Count | As % |
|---|---|---|
| Members of the public | 1 | 100% |
Mendeley readers
Geographical breakdown
| Country | Count | As % |
|---|---|---|
| Unknown | 7 | 100% |
Demographic breakdown
| Readers by professional status | Count | As % |
|---|---|---|
| Researcher | 2 | 29% |
| Student > Bachelor | 1 | 14% |
| Other | 1 | 14% |
| Student > Master | 1 | 14% |
| Student > Ph. D. Student | 1 | 14% |
| Other | 0 | 0% |
| Unknown | 1 | 14% |
| Readers by discipline | Count | As % |
|---|---|---|
| Biochemistry, Genetics and Molecular Biology | 3 | 43% |
| Agricultural and Biological Sciences | 2 | 29% |
| Neuroscience | 1 | 14% |
| Unknown | 1 | 14% |