Ribosome Affinity Purification Service

Overview Application Why Us FAQs Related Services

Are you currently struggling to identify truly translated mRNA targets, uncertain about the functional impact of mRNA isoforms, or needing to pinpoint changes in protein synthesis under specific conditions? At Creative Biolabs, our ribosome affinity purification (RAP) service helps you accurately identify and quantify actively translated mRNAs, enabling you to gain deeper insights into gene regulation at the translational level through our optimized RAP protocols, advanced affinity purification strategies, and comprehensive downstream analysis capabilities.

Project workflow

Our RAP service is meticulously engineered to provide an encompassing and completely transparent journey. We'll be your dedicated companion, guiding you seamlessly from the moment you submit your initial samples right up to the point where you have actionable data in hand. Below are the pivotal steps that make up this process:

Consultation and Experimental Design

A thorough discussion of your project goals, selection of appropriate tagged ribosomal proteins, optimization of cell culture and lysis conditions, and planning for downstream analysis.
Cell Culture and Lysis

Culturing cells under specified conditions, treatment with translation inhibitors to stabilize ribosome-mRNA complexes, and gentle cell lysis to preserve polysome integrity. For tissue samples, suitable homogenization and processing protocols are implemented.
Affinity Purification of Ribosomes

Immunoprecipitation of tagged ribosomes using high-specificity affinity reagents. This step is meticulously optimized to maximize yield and purity while minimizing non-specific binding.
RNA Isolation and Quality Control

Extraction of RNA from the purified ribosome complexes. Rigorous quality control measures, including assessment of RNA integrity and quantity, and DNase treatment to remove any genomic DNA contamination.
Downstream Analysis

Preparation of libraries for NGS, RT-qPCR analysis of specific targets, or other requested analyses. Bioinformatic processing of sequencing data, including mapping, quantification, and differential expression analysis.

Application Areas

Cell-Type-Specific Translational Profiling

In complex tissues with various cell types, RAP Service enables targeted ribosome purification from specific populations. This is done by expressing ribosome-binding affinity tags in a cell-type-specific way. Analyzing the mRNAs on these isolated ribosomes reveals each cell type's unique translational profile, uncovering actively translated genes and their regulatory mechanisms.

Developmental Biology

During development, cells show dynamic gene expression and protein synthesis changes. RAP Service tracks translational alterations at different stages. Isolating ribosomes from developing tissues/organs helps identify mRNAs preferentially translated in processes like organogenesis, cell differentiation, and morphogenesis, aiding the understanding of developmental transitions and translational control.

Disease Research & Biomarker Discovery

Abnormal protein synthesis is common in diseases. RAP Service compares translational profiles between healthy and diseased cells/tissues. Identifying differentially translated mRNAs in disease states points to potential therapeutic targets and disease-specific biomarkers.

Stress Response Analysis

Cells adjust gene expression at the translational level under environmental stresses like heat shock, oxidative stress, or nutrient deprivation. RAP Service captures translational changes under stress. Isolating ribosomes from stressed cells and analyzing associated mRNAs identifies up or down-regulated genes, revealing cellular stress-coping strategies and molecular mechanisms.

Neuroscience

The brain's complexity, with diverse neuronal types and circuits, makes RAP Service valuable. It studies translational regulation in specific neuronal populations, aiding the understanding of neural functions, synaptic plasticity, and neurological disorders. Isolating ribosomes from different brain regions or neuronal subtypes reveals genes critical for neural development, memory formation, and neurodegenerative disease pathogenesis.

Why Choose Us?

Creative Biolabs is steadfastly dedicated to offering state-of-the-art RAP services that yield highly reliable and profoundly insightful data. Our entire methodology rests on a solid bedrock of scientific prowess and an unwavering commitment to top-notch quality.

High Specificity and Purity

We utilize optimized protocols, including the potential use of advanced reagents for tagged ribosome immunoprecipitation, to ensure highly specific capture of translating ribosomes and minimize co-purification of non-ribosomal contaminants.

Expertise with Diverse Sample Types

Our team has experience processing a variety of biological samples, including mammalian cells, yeast, and bacteria, adapting protocols to suit specific experimental systems.

Comprehensive Downstream Options

We offer a suite of downstream analysis services, from RT-qPCR to full RNA sequencing and bioinformatics, providing a seamless workflow from sample to publication-ready data.

Customizable Service

Creative Biolabs offers flexible service options, from providing purified ribosome-associated RNA to full end-to-end analysis, tailored to your specific needs and budget.

Transparent Communication

We ensure clear and consistent communication at every stage of your project, delivering regular updates and comprehensive reports.

FAQs

Q: How does RAP compare to ribosome profiling?

A: RAP isolates full-length mRNAs associated with ribosomes, giving you information about which mRNAs are being translated. Ribo-seq, on the other hand, sequences only the ribosome-protected mRNA fragments, providing positional information about ribosomes on the mRNA. Both are formidable, the best choice depends on your research question. We can help you decide which combination is best for your project.

Q: Can RAP be used to study non-coding RNAs?

A: RAP is primarily designed to capture mRNAs actively engaged with ribosomes. While some non-coding RNAs might associate with ribosomes in regulatory roles, our protocols are optimized to enrich for translating mRNAs.

Q: What species are compatible with the RAP service offered by Creative Biolabs?

A: Our capabilities allow us to isolate ribosomes from an extensive array of species. This includes human, yeast, bacterial, and plant samples. Should you require more detailed information regarding specific species, please get in touch with us.

Q: What quality control measures do you have in place for the RAP service?

A: We implement multiple QC steps, including validation of tagged protein expression, assessment of RNA integrity post-purification, and checks for genomic DNA contamination.

Q: What advantages does utilizing your RAP service offer compared to handling the task in-house?

A: Our service offers access to optimized protocols, specialized reagents, experienced personnel, and dedicated instrumentation, potentially saving you significant time and resources in establishing and troubleshooting the technique.

Related Services

Ready to see how we can streamline your research? Get in touch with us for further details.

For Research Use Only. We do not provide direct services or products for patients.

Services

Online Inquiry