Ribosome Affinity Purification (RAP) Service
Ribosomes play an important role in protein synthesis. Abnormal synthesis of ribosomes is associated with various diseases, such as hematologic diseases and cancers. In recent years, ribosome analysis has gradually become a hotspot. As a leading service provider of ribosome purification, Creative Biolabs will provide customized ribosome affinity purification (RAP) and ribosome analysis services according to your requirements.
Background
Ribosomes are composed of large and small subunits, each of which is an RNA/protein complex. In the process of biological development, the morphology of ribosomes has changed, and the structural changes have also provided a new direction for the study of interacting proteins. To probe this protein interaction, the RAP method was proposed, which can identify hundreds of ribosome-associated proteins, as well as a variety of RNA and protein modifying enzymes. Currently, this technique plays an important role in ribosome analysis.
RAP is considered to be a method to rapidly purify ribosomes for further relevant information. The method relies on the expression of tagged versions of specific ribosomal proteins (RPs), and IgG-coupled spherical microbeads (matrix) efficiently recover ribosomes from cell extracts with the assistance of the tagged proteins. It is worth noting that this method plays an important role in analyzing all the information associated with the ribosome. Most importantly, RPs and their modifications can be mapped using mass spectrometry. This approach provides a promising direction for further exploration of gene expression patterns.
Fig. 1 Inhibition of RP translation prevents reduction of long mRNA translation during mGluR-LTD.¹
Ribosomes consist of ribosomal proteins RPs and ribosomal RNA (rRNA), with the key to the success of Ribosome Affinity Purification (RAP) lying in the careful selection of an RP suitable for affinity labeling. Importantly, fusion labels do not interfere with the function of RPs. Affinity-labeled ribosomes are captured using IgG-conjugated spherical microspheres and can be efficiently released from the matrix with proper handling. RAP offers significant advantages, including the rapid acquisition of purified ribosomes, prevention of contamination from impurities like lipid rafts and pseudopolysomes, and the use of sucrose-free extraction solutions, which is particularly advantageous for subsequent mass spectrometry analysis. Additionally, RNA isolation is straightforward, facilitating further experimental procedures. However, RAP also has some limitations; it provides only a broad overview of ribosome-related information and cannot distinguish whether the mRNA is bound to one or multiple ribosomes, nor does it indicate the extent to which the associated information has been translated.
Our Service
At Creative Biolabs, our Ribosome Affinity Purification (RAP) service offers a comprehensive, customizable platform for ribosome isolation that accommodates diverse biological samples, including bacteria, yeast, and mammalian cells. By leveraging advanced affinity-tagging methods, we ensure the high-purity isolation of ribosomes, suitable for various downstream applications such as proteomics, RNA sequencing, and structural biology. Our platform supports multiple affinity tags, while employing optimized buffer systems to maintain the integrity and functionality of ribosomes throughout the process. This service is adaptable to a broad range of species, making it ideal for researchers seeking specific ribosomal components for detailed analysis. Additionally, the isolated ribosomes are compatible with a wide array of experimental applications, allowing researchers to perform mass spectrometry, ribosome profiling, and more.
Highlights
- High Yield & Purity
Our protocols ensure high-quality ribosome samples, minimizing contamination and maximizing experimental reproducibility.
- Comprehensive Support
From consultation to final delivery, our scientific experts offer end-to-end guidance, ensuring optimal outcomes for your ribosome research.
- Custom Solutions
Tailored purification protocols to meet diverse research needs, from small-scale exploratory studies to large-scale ribosome profiling projects.
- Cost-Effective & Efficient
Short turnaround times and competitive pricing without compromising quality.
FAQs
Q1: What species are supported for the RAP service at Creative Biolabs?
A: We can isolate ribosomes from a wide range of species, including human, bacterial, yeast, and plant samples. Please contact us for more details about specific species.
Q2: What downstream applications can the purified ribosomes be used for?
A: Purified ribosomes are suitable for proteomic studies, RNA sequencing, ribosome profiling, and interaction studies with translation factors and regulatory proteins.
Q3: How does RAP compare with traditional methods of ribosome isolation?
A: RAP provides a higher level of specificity and yield compared to conventional centrifugation methods. The use of affinity tags significantly enhances the purification process, reducing contamination and ensuring more targeted results.
Q4: What are the key advantages of using RAP for ribosome isolation?
A: RAP offers quicker isolation times, high specificity, and the ability to isolate intact ribosomes suitable for downstream analyses, making it a preferred method for ribosome research.
Resource
Ribosome Analysis - Creative Biolabs
Ribosome Analysis - Creative BiolabsAfter decades of accumulation, we have made great progress and accumulated rich experience in the field of ribosome purification and analysis. With our professional team of scientists and rich experience, Creative Biolabs is committed to providing optimal RAP service to customers around the world. Please contact us for more details.
Reference
- Seo, Sang S., et al. "Excess ribosomal protein production unbalances translation in a model of Fragile X Syndrome." Nature Communications 13.1 (2022): 3236.
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