MRM Ribosomal Proteomic Analysis Service
Are you currently facing challenges in precisely quantifying low-abundance ribosomal proteins (RPs), validating disease-specific biomarkers, or unraveling complex translational regulatory mechanisms? Our multiple reaction monitoring (MRM) ribosomal proteomic analysis service helps you gain unprecedented insights into cellular function and accelerate your research through advanced, highly sensitive, and reproducible targeted protein quantification using state-of-the-art mass spectrometry technology.
A Brief Introduction to MRM
Ribosomes, the cellular machinery responsible for protein synthesis, are complex ribonucleoprotein complexes comprising ribosomal RNA and numerous RPs. Variations in RP composition, expression levels, and post-translational modifications (PTMs) are increasingly recognized for their critical roles in regulating gene expression and influencing cellular physiology and disease, including cancer and neurodegeneration. MRM, a highly sensitive and specific mass spectrometry-based technique, has emerged as the "gold standard" for targeted quantitative proteomics. By precisely quantifying specific peptides, MRM enables robust and reproducible analysis of ribosomal proteomes, offering unprecedented insights into their dynamic states. This targeted approach overcomes limitations of traditional methods, providing accurate quantification of low-abundance proteins and PTMs, which is essential for understanding ribosomal function and its implications in health and disease.
Fig.1 Schematic overview of targeted quantitative proteomics methods.1
Simplified Workflow
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Sample Preparation & Digestion
RPs samples undergo rigorous quality control, followed by enzymatic digestion into peptides. This crucial step prepares the proteins for mass spectrometry analysis, ensuring optimal peptide recovery and compatibility with our instruments. -
MRM Method Development & Optimization
Based on your target RPs, our experts design and optimize MRM assays. This involves selecting unique peptides and their characteristic fragment ions and fine-tuning mass spectrometer parameters to achieve maximal sensitivity and specificity for your specific targets.
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LC-MS/MS Data Acquisition
Prepared peptide samples are introduced into high-performance liquid chromatography (LC) systems. This stage involves the precise and highly selective quantification of your target ribosomal peptides based on the optimized MRM transitions.
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Data Processing & Quantification
Raw mass spectrometry data is rigorously processed using advanced bioinformatics tools. This includes peak integration, normalization, and relative or absolute quantification of your target RPs, often utilizing stable isotope-labeled internal standards for precise absolute quantification.
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Bioinformatics Analysis & InterpretationBeyond raw numbers, our team performs in-depth bioinformatics analysis, including statistical assessment of differential protein expression, functional annotation, pathway analysis, and protein-protein interaction network mapping.
Final Outputs
- Comprehensive Quantitative Protein Reports
Detailed reports including quantification data for all targeted RPs, statistical analysis, and visual representations, such as volcano plots, heatmaps.
- Raw Mass Spectrometry Data Files
All original LC-MS/MS raw data for your records and further in-house analysis, if desired.
- Detailed Bioinformatics Analysis
Functional enrichment analysis, pathway mapping, and any custom analyses requested, providing a deeper understanding of the biological implications of your ribosomal proteomic changes.
What Makes Us the Preferred Choice?
- Superior Analytical Performance
The core of our service is built upon the inherent strengths of MRM technology, which we optimize to its fullest potential.
- Highest Accuracy
Through highly selective ion monitoring, we achieve exceptional accuracy in protein quantification, minimizing interference from complex biological matrices.
- Unmatched Sensitivity
Our optimized methods allow for the detection and quantification of RPs and their PTMs even at femtomole levels, critical for discovering subtle but significant changes.
- Exceptional Reproducibility
Rigorous quality control, standardized protocols, and instrument stability ensure highly reproducible results across experiments and batches, providing reliable data for your critical decisions.
- Broad Dynamic Range
We effectively cover a wide range of protein abundances, enabling the quantification of both highly expressed and low-abundance ribosomal components.
Applications
- Translational Regulation Studies
Quantifying changes in RPs stoichiometry and modifications to understand how translation is regulated under different physiological or pathological conditions.
- Ribosomopathy Research
Identifying and quantifying specific RPs dysregulations linked to inherited or acquired ribosomal disorders, aiding in disease mechanism elucidation and therapeutic development.
- Verification of Untargeted Proteomics Data
Providing high-confidence, quantitative validation for RPs candidates identified through broader, untargeted proteomic screens, such as label-free or TMT-based approaches.
- Cellular Stress Response Profiling
Quantifying changes in RPs composition and modifications in response to various cellular stressors, such as nutrient deprivation, oxidative stress, or environmental toxins.
FAQs
Q: How sensitive is the MRM ribosomal proteomic analysis service, especially for low-abundance RPs or PTMs?
A: MRM is renowned for its exceptional sensitivity. By selectively monitoring specific precursor-to-fragment ion transitions, our service significantly reduces background noise, allowing for the reliable detection and quantification of RPs and their PTMs even at femtomole concentrations. This high sensitivity is critical for uncovering subtle yet significant biological changes.
Q: Can your service differentiate between highly homologous RPs?
A: Absolutely. One of the key strengths of MRM is its unparalleled specificity. We design and optimize unique peptide transitions for each target, enabling us to distinguish and quantify highly homologous RPs that might be challenging to resolve with other methods. This is particularly valuable for studying ribosomal heterogeneity.
Q: What kind of bioinformatics support is included with the MRM ribosomal proteomic analysis service?
A: Our service includes comprehensive bioinformatics analysis. This ranges from raw data processing and quality assessment to advanced statistical analysis for differential expression, functional annotation, clustering analysis, and protein-protein interaction network mapping. We ensure you receive interpretable and actionable biological insights from your quantitative data.
Creative Biolabs is dedicated to advancing your scientific endeavors with our leading MRM ribosomal proteomic analysis service. By combining cutting-edge mass spectrometry technology with deep scientific expertise, we deliver highly accurate, sensitive, and reproducible quantitative data, empowering your research into ribosomal biology. Ready to unlock the full potential of your ribosomal proteomics research? Connect with our expert team 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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