| Item type |
文献 / Documents(1) |
| 公開日 |
2024-10-11 |
| アクセス権 |
|
|
アクセス権 |
open access |
|
アクセス権URI |
http://purl.org/coar/access_right/c_abf2 |
| 資源タイプ |
|
|
資源タイプ識別子 |
http://purl.org/coar/resource_type/c_6501 |
|
資源タイプ |
journal article |
| 出版社版DOI |
|
|
|
関連識別子 |
https://doi.org/10.21769/BioProtoc.4106 |
|
|
関連名称 |
10.21769/BioProtoc.4106 |
| 出版タイプ |
|
|
出版タイプ |
VoR |
|
出版タイプResource |
http://purl.org/coar/version/c_970fb48d4fbd8a85 |
| タイトル |
|
|
タイトル |
Efficient and Rapid Analysis of Polysomes and Ribosomal Subunits in Cells and Tissues Using Ribo Mega-SEC |
| 著者 |
𠮷川, 治孝
Sundaramoorthy, Ramasubramanian
Mariyappa, Daniel
Jiang, Hao
Lamond, Angus I
|
| 抄録 |
|
|
内容記述 |
Polysome profile analysis is a popular method for separating polysomes and ribosomal subunits and is typically achieved using a sucrose density gradient (SDG). This has remained the gold standard method sinceribosomes were first discovered; however, this method is time-consuming and requires multiple steps from making the gradient and long ultracentrifugation to collecting and analyzing the fractions. Each of these steps in the SDG workflow can introduce potential technical variation that affects the reproducibility of gradient profiles between samples. To address these limitations, we have developed a flexible, alternative approach for analyzing polysomes and ribosomal subunits based on size-exclusion chromatography (SEC), termed ‘Ribo Mega-SEC.’ In comparison with the SDG method, Ribo Mega-SEC involves a single step using ultra-high-performance liquid chromatography (uHPLC). The entire workflow, from injecting the lysate to collecting the fractions, can be performed in as little as 15 min, with high reproducibility. By varying the pore size of the SEC column, polysomes and ribosomal subunits can be separated using extracts from either human or mouse cultured cell lines or from tissue samples, Drosophila embryos, or budding yeast. The resulting separated fractions are suitable for analysis using a wide range of subsequent analytical techniques including mass spectrometry (MS)-based proteomics, RNA-Seq, electron microscopy (EM), and multiple biochemical assays. |
| キーワード |
|
|
主題 |
Polysome |
| キーワード |
|
|
主題 |
Ribosome |
| キーワード |
|
|
主題 |
Polysome profile |
| キーワード |
|
|
主題 |
Size-exclusion chromatography |
| キーワード |
|
|
主題 |
Sucrose density gradient |
| キーワード |
|
|
主題 |
mRNA translation |
| キーワード |
|
|
主題 |
Protein synthesis |
| 書誌情報 |
en : Bio-protocol
巻 11,
号 15,
p. e4106,
発行日 2021-08-05
|
| 収録物ID |
|
|
収録物識別子タイプ |
EISSN |
|
収録物識別子 |
23318325 |
| 出版者 |
|
|
出版者 |
Bio-protocol |
| 権利情報 |
|
|
権利情報 |
This article is distributed under the terms of the Creative Commons Attribution License. |
| EID |
|
|
識別子 |
381466 |
| 言語 |
|
|
言語 |
eng |
bioprotoc_11_15_e4106.pdf (654 KB)
.png)
