PRM Ribosomal Proteomic Analysis Service
Are you currently facing challenges in accurately quantifying low-abundance ribosomal proteins (RPs), identifying post-translational modifications (PTMs) on ribosomes, or validating crucial ribosomal biomarkers in complex biological samples? Our parallel reaction monitoring (PRM) ribosomal proteomic analysis service helps you gain unparalleled insights into ribosomal function and dynamics, enabling precise quantification of targeted RPs and their modifications through cutting-edge PRM mass spectrometry (MS).
Overview of PRM
The ribosome, the cell's protein synthesis machinery, is far more dynamic and heterogeneous than previously thought. Variations in RP composition and PTMs are emerging as critical regulators of gene expression, influencing everything from cell differentiation to disease pathology, including cancer and neurological disorders. Traditional proteomic approaches often struggle with the comprehensive and quantitative analysis of RPs due to their high abundance, complex modifications, and the challenges of isolating pure ribosomal fractions. PRM technology, leveraging high-resolution MS, offers a powerful solution, enabling the precise and sensitive quantification of targeted RPs and their specific modifications, opening new avenues for understanding translational control and identifying novel disease biomarkers.
Fig.1 Schematic overview of targeted quantitative proteomics methods.1
Main Process
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Sample Preparation and Digestion
Client-provided ribosomal samples undergo rigorous quality control. Proteins are then extracted and enzymatically cleaved into peptides. This step is optimized to ensure maximal recovery and reproducibility of ribosomal peptides. -
Peptide Enrichment
For low-abundance peptides or specific PTMs, enrichment strategies may be employed to enhance detection sensitivity.
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PRM Assay Development & Optimization
Based on your target list, proteotypic peptides are selected, and PRM assays are developed and optimized on our high-resolution MS platforms. This involves establishing optimal collision energies and monitoring transitions for each target peptide to ensure specificity and sensitivity.
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High-Resolution LC-MS/MS Analysis
Prepared peptide samples are subjected to liquid chromatography (LC) for separation, followed by PRM MS using state-of-the-art instruments. This step ensures precise and simultaneous quantification of multiple target peptides.
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Data Acquisition and Bioinformatics Analysis
Raw MS data are acquired and then processed using advanced bioinformatics software. This involves rigorous peak integration, quality control, and quantitative analysis to determine the relative or absolute abundance of your target RPs and PTMs across samples.
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Statistical Analysis and Report GenerationComprehensive statistical analyses are performed to identify significant changes in RP levels or modifications between experimental groups. A detailed report is generated, including all raw data, processed quantitative results, statistical findings, and biological interpretations.
Why Choose Creative Biolabs?
Creative Biolabs is a leader in targeted proteomics, offering a PRM ribosomal proteomic analysis service that stands apart for its unparalleled precision, reproducibility, and expert-driven insights. Our commitment to cutting-edge technology and scientific excellence ensures your ribosomal research yields reliable and impactful results. We leverage state-of-the-art MS platforms, including high-resolution Orbitrap systems with advanced acquisition modes, providing attomole-level sensitivity and a broad dynamic range essential for the intricate world of ribosomal proteomics. Our experienced team of scientists specializes in optimizing targeted proteomic workflows, ensuring robust data even from challenging ribosomal preparations. Our expertise translates into superior data quality, faster turnaround times, and a deeper understanding of your RP dynamics.
Applications
- Translational Control Studies
Investigating how changes in RP composition or modifications influence protein synthesis efficiency and specificity.
- Antibiotic Resistance Mechanisms
Elucidating how ribosomal modifications contribute to bacterial resistance to antibiotics aids in the development of new antibacterial strategies.
- Cellular Stress Response Analysis
Elucidating the functional implications of ribosomal modifications in cellular adaptation and stress response mechanisms.
- PTMs Profiling
Precise quantification of specific ribosomal PTMs to uncover their functional significance in cellular processes.
- Host-Pathogen Interaction Studies
Investigating how pathogens manipulate host ribosomes and how host cells alter ribosomal composition in response to infection.
FAQs
Q: What types of RPs or modifications can your PRM service quantify?
A: Our PRM ribosomal proteomic analysis service is highly versatile, capable of quantifying a wide range of RPs and various PTMs, including phosphorylation, methylation, acetylation, and ubiquitination, provided appropriate assays can be developed. Share your specific targets with us, and we can discuss the feasibility for your unique project.
Q: How does PRM compare to other quantitative proteomics methods for ribosomal analysis?
A: PRM offers superior specificity, sensitivity, and reproducibility for targeted quantification compared to discovery-based methods. While DIA provides comprehensive coverage, PRM excels when you need highly precise and confident quantification of a predefined set of ribosomal targets. Contact us to determine the best approach for your research goals.
Q: What are the typical sample requirements for RP analysis using PRM?
A: We generally recommend purified ribosomal fractions or highly enriched ribosomal pellets to minimize interference from highly abundant non-RPs. Specific quantities vary by sample type, such as cells, tissues, and biofluids, but our team can provide detailed guidelines and advice on optimal preparation methods to ensure successful analysis. Reach out to discuss your specific sample needs!
Q: Can your PRM service help in understanding antibiotic resistance mechanisms?
A: Absolutely. Ribosomes are key targets for many antibiotics, and changes in RP composition or modifications can confer resistance. Our PRM service can precisely quantify these changes, helping you identify and validate specific ribosomal alterations linked to antibiotic resistance, accelerating your research in infectious diseases.
From quantifying subtle changes in RP abundance to identifying critical PTMs, our targeted approach delivers the accurate, reproducible data you need to drive your discoveries. We are committed to accelerating your research with our cutting-edge technology and scientific excellence. Partner with Creative Biolabs today!
Reference
- Ryu, Joohyun, and Stefani N. Thomas. "Quantitative mass spectrometry-based proteomics for biomarker development in ovarian cancer." Molecules 26.9 (2021): 2674. DOI: 10.3390/molecules26092674. Distributed under Open Access license CC BY 4.0, without modification.
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