TMT Ribosomal Proteomic Quantification Service
Are you struggling to identify actively translated proteins that are pivotal for your research? Our TMT (tandem mass tag) ribosomal proteomics quantification service helps you precisely measure active protein synthesis and identify changes in ribosome occupancy across multiple conditions. Through advanced TMT labeling coupled with ribosome footprinting and state-of-the-art LC-MS/MS analysis, we empower you to accelerate your discovery of novel therapeutic targets and gain deeper insights into complex biological processes.
Introduction to TMT Ribosomal Proteomics Quantification
TMT ribosomal proteomics quantification is a sophisticated technique that measures active protein synthesis by quantifying ribosome-associated mRNA. It combines ribosome profiling with TMT isobaric labeling for multiplexed quantitative proteomics. This approach allows researchers to gain a dynamic snapshot of the translatome, revealing how protein synthesis is regulated under various conditions, and complementing transcriptomic data by showing which mRNAs are actively being translated and at what levels.
Fig.1 Workflow of TMT labeling-based proteome profiling in B.napus seeds.1
Workflow
Our TMT ribosomal proteomics quantification workflow is designed for robustness and clarity, ensuring high-quality data and actionable insights.
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Sample Quality Control (QC) & Preparation
Initial assessment of sample integrity. Cells or tissues are lysed under conditions that preserve ribosome-mRNA complexes. -
Protein Extraction
Ribosomal proteins are extracted from cells, tissues, or biofluids and enriched using specialized methods to ensure high purity and yield.
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TMT Labeling & Sample Multiplexing
Peptides from each sample are labeled with distinct TMT reagents, allowing for simultaneous analysis of multiple samples. Labeled samples are then combined.
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Peptide Fractionation & LC-MS/MS Analysis
The pooled, TMT-labeled peptide sample is fractionated by high-pH reversed-phase chromatography to reduce complexity. Each fraction is then analyzed by high-resolution Orbitrap LC-MS/MS.
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Bioinformatics Analysis & Data InterpretationRaw data is processed to identify and quantify TMT reporter ions, mapping peptides to proteins and determining relative changes in ribosome occupancy or protein synthesis across samples.
Final Deliverables
- A detailed project report summarizing the methodology, QC parameters, identified proteins, and quantitative data on differential ribosome occupancy or protein synthesis.
- Raw data files and processed data tables including peptide and protein identifications, quantitation values, and statistical analysis.
- Bioinformatics analysis results may include pathway analysis, Gene Ontology (GO) enrichment, and visualization plots.
Why Choose Creative Biolabs?
- Expert Insights
Creative Biolabs specializes in TMT ribosomal proteomics, providing high-fidelity translational regulation insights. Our expert team drives your research with deep expertise and nuanced analysis, ensuring actionable, accurate results.
- Advanced TMT Tech
Our cutting-edge TMT reagents enable superior multiplexing and quantification accuracy. This advanced technology allows simultaneous analysis of multiple samples, capturing even subtle protein changes with exceptional precision.
- Comprehensive Analysis
Combining optimized ribosome footprinting with sensitive LC-MS/MS, we achieve comprehensive proteome coverage. This approach efficiently captures ribosome-protected mRNA fragments, including low-abundance proteins, for detailed translational regulation studies.
- Tailored Bioinformatics
Our skilled bioinformaticians customize data analysis using advanced tools and algorithms. They identify differentially expressed proteins and regulatory mechanisms, translating complex data into clear, research-relevant conclusions tailored to your objectives.
- Strict QC Measures
Rigorous quality control ensures reliable, reproducible results. Strict protocols, including instrument calibration and data validation, guarantee accuracy at every step, providing confidence in your research findings.
Applications
Our TMT ribosomal proteomics quantification service offers profound insights across a wide range of research and development areas:
- Drug Discovery and Development
Leverage TMT ribosomal proteomics to identify novel therapeutic targets by detecting proteins with dysregulated translation in disease. Elucidate drug mechanisms through direct observation of protein synthesis rate changes, assess efficacy, and uncover off-target translational effects for safer, more precise therapies.
- Biomarker Discovery
Pinpoint translational biomarkers by analyzing active protein synthesis profiles correlated with disease states, progression, or treatment responses. Identify markers of translational stress or reprogramming to enhance diagnostics and therapeutic monitoring in diverse clinical contexts.
- Disease Mechanism Research
Investigate translational control in complex diseases like cancer, neurodegeneration, and metabolic syndromes. Uncover disease-specific ribosome dysfunction or protein synthesis fidelity alterations to reveal new pathogenic pathways and intervention opportunities.
- Fundamental Biological Insights
Study translational regulation under cellular stressors and across developmental stages. Map cell-type-specific translatomes to decode how protein synthesis dynamics govern cellular behavior and adaptation.
- Systems Biology Integration
Combine translatomic data with genomic, transcriptomic, and proteomic datasets for holistic cellular process modeling. Validate RNA-seq findings by correlating mRNA abundance changes with actual protein synthesis outputs, strengthening multi-omics insights.
- Biotechnology Optimization
Improve recombinant protein production by resolving translational bottlenecks in expression systems. Monitor and enhance translational efficiency of engineered metabolic pathways to accelerate bioprocess development and industrial biotechnology applications.
FAQs
Q: What are the main advantages of TMT ribosomal proteomics over standard total proteomics?
A: TMT ribosomal proteomics specifically measures actively synthesizing proteins by targeting ribosome-bound mRNAs or nascent peptides. This provides a more dynamic view of the cell's response to stimuli compared to total proteomics, which reflects steady-state protein levels.
Q: What types of samples are suitable for this service?
A: We can process a variety of sample types, including cultured cells, tissues from animal models, and patient-derived samples. The key is proper sample collection and preservation to maintain the integrity of ribosome-mRNA complexes.
Q: How does TMT ribosomal proteomics compare to ribosome profiling by sequencing (Ribo-Seq)?
A: Ribo-Seq identifies and quantifies mRNA fragments protected by ribosomes, providing insights into ribosome occupancy at the RNA level. TMT ribosomal proteomics, as offered by Creative Biolabs, typically quantifies the protein output associated with these ribosomes, directly measuring protein synthesis or ribosome composition using mass spectrometry.
Q: What level of bioinformatics analysis is included?
A: Our standard service includes peptide/protein identification, TMT reporter ion quantification, statistical analysis to identify differentially synthesized proteins, and basic pathway/GO enrichment analysis. We also offer customizable bioinformatics solutions to address specific research questions.
At Creative Biolabs, our TMT ribosomal proteomics quantification service offers unparalleled insight into the dynamic world of protein synthesis. We are committed to providing you with high-quality, actionable data and expert support to accelerate your research and discovery. Ready to explore how TMT ribosomal proteomics can advance your project? Our expert team is here to assist you.
Reference
- Zhou, Zhongjing, et al. "Tandem Mass Tag-Based Quantitative Proteomics Reveals Implication of a Late Embryogenesis Abundant Protein (BnLEA57) in Seed Oil Accumulation in Brassica napus L." Frontiers in Plant Science 13 (2022): 907244. Distributed under Open Access license CC BY 4.0, without modification.
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