RNA binding protein (RBP) is a powerful and widespread class of regulatory factors, accounting for about 5% to 10% of all proteins encoded in cells. Currently, except for a few RNAs that function as nucleases alone, most RNAs function by binding to proteins to form RNA-protein complexes. With the advent of the genetic era, RNA-related research has become an important part of biology. It has been widely studied in the fields of tumor, chronic diseases such as diabetes, enteritis, etc. However, the mechanism of RNA is still poorly understood, and various newly discovered IncRNA, circRNA and protein interactions need to be further investigated and discovered.
On the one hand, RBP is involved in several post-transcriptional regulatory processes such as RNA synthesis, variable splicing, modification, translocation, and intracellular localization, regulating biological processes such as mRNA stability, lncRNA activity, and miRNA silencing complex formation; on the other hand, RNA also regulates the activity, localization, or interactions with other proteins of these proteins, thus affecting various RBP-mediated cellular events. The interaction between RNA and proteins is very important for maintaining cellular homeostasis, and disruption of this interaction will lead to cellular dysfunction and related diseases.
The current methods to study RNA-protein interactions are divided into two major categories: one is to find the protein that binds to the RNA of interest; the other is to find the RNA that binds to the RNA of interest. rna pull-down belongs to the former, which tags the RNA of interest by means of in vitro transcription, binds the RNA to the resin support through this tag, then adds the sample protein to bind to the RNA, washes and elutes to obtain the protein that binds to the RNA. RNA pull-down is one of the main techniques to detect the interaction between RNA-binding proteins and their target RNAs, and it is the main means to study the interaction between IncRNA, circRNA and proteins in recent years.
RNA pull down uses in vitro transcription to label biotin RNA probes, which are then incubated with cytosolic protein extracts to form RNA-protein complexes. This complex can be bound to streptavidin-labeled magnetic beads, thereby separating from other components in the incubation solution. After the complex is eluted, whether a specific RNA-binding protein interacts with RNA can be detected by Western Blot, and unknown interacting proteins can also be identified by mass spectrometry.
Creative Proteomics has established a sophisticated molecular experimental platform. All you need to do is provide the target protein antibody (available for Western Blot) and its instructions, the cell sample to be tested, the target gene, and the target protein information. Our skilled technical team will then provide you with RNA Pull-Down Result figure, raw data, result figure (including Input, positive and negative control results), and standard experiment report (including detailed materials and methods).
RNA Pull Down Service Workflow
Animal tissue sample: >100mg
Plant tissues >5g
Creative Proteomics has independent laboratories for proteomics, interactomics, and molecular cells. The laboratories are equipped with large precision instruments such as liquid chromatography-mass spectrometry (LC-MS), gas chromatography-mass spectrometry (GC-MS), isoelectric focusing instrument and various specialized analytical testing equipment. We realize the traceability of data results and ensure the accuracy of experimental results.
Creative Proteomics provides high quality technical services to biopharmaceutical companies, universities, research institutions, hospitals and individuals. We have assisted our clients in publishing papers in high level journals such as Nature methods, Journal of Hematology & Oncology, etc.