知无不“研”| 一文读懂RNA pull down - 知乎 - 知乎专栏
2020年7月30日 · RNA pull-down作为研究RNA与蛋白互作的核心技术，已成为研究焦点。 RNA pull down是使用 体外转录法 标记生物素RNA探针，然后与胞浆蛋白提取液孵育，形成 RNA-蛋白质 复合物。 该复合物可与链霉亲和素标记的磁珠结合，从而与孵育液中的其他成分分离。 复合物洗脱后，通过WB实验检测特定的RNA结合蛋白是否与RNA相互作用，或者结合MS筛选RNA …
Three micrograms of biotinylated RNA was heated to 90°C for 2 minutes, put on ice for 2 minutes, supplied with RNA structure buffer (10 mM Tris pH 7, 0.1 M KCl, 10 mM MgCl2), and then shifted to room temperature (RT) for 20 minutes to allow proper secondary structure formation.
RNA Pull Down: Principles, Probe Design, and Data Analysis
The basic principle of RNA Pull Down involves the isolation of RNA-binding proteins (RBPs) that interact with a specific RNA of interest. This technique utilizes biotinylated RNA probes that are complementary to the target RNA sequence.
Methods for Detecting Protein–RNA Interactions | Thermo …
We offer the Thermo Scientific Pierce Magnetic RNA-Protein Pull-Down Kit, which provides reagents to efficiently enrich RNA-binding proteins (RBPs) using RNA end-labeled with desthiobiotin and streptavidin magnetic beads.
Methods to study RNA–protein interactions - Nature
2019年2月25日 · Recent progress has expanded the number of methods for studying RNA–protein interactions in two general categories: approaches that characterize proteins bound to an RNA of interest...
RNA pulldown protocol for in vitro detection and identification of RNA ...
Here we describe a detailed RNA pulldown protocol for the in vitro detection of proteins specifically interacting with a long RNA molecule. Recent advances in genomics have revealed that cells encode thousands of noncoding RNA molecules that do not require translation to proteins to perform their biological roles.
Pierce™ Magnetic RNA-Protein Pull-Down Kit - Thermo Fisher Scientific
The Magnetic RNA-Protein Pull-Down Kit provides reagents to efficiently enrich RNA Binding Proteins (RBPs) using RNA end-labeled with desthiobiotin and streptavidin magnetic beads. The complete kit contains sufficient reagents for 20 RNA labeling reactions and …
RNA Pull-down Procedure to Identify RNA Targets of a Long Non …
2018年4月10日 · Coupled with high throughput RNA sequencing, this RNA pull-down protocol can provide the whole RNA interactome of a lncRNA of interest. Long non-coding RNA (lncRNA), which are sequences of more than 200 nucleotides without a defined reading frame, belong to the regulatory non-coding RNA's family.
cultured cells that have to preserve the network between all molecular partners. Coupled with high throughput RNA sequencing, this RNA pull-down protocol can provide the whole RNA interactome of a lncRNA of interest. Video Link The video component of this article can be found at https://www.jove.com/video/57379/ Introduction
Enzymolysis-based RNA pull-down identifies YTHDC2 as an …
2023年10月31日 · Zhu et al. report an enzymolysis-based RNA pull-down method using IdeS and identify YTHDC2 as an m6A-modified IFN-β mRNA-binding protein that inhibits IFN-β expression and antiviral innate response through recruiting ISG20 in cells and mice.
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