Western Blotting (WB) Service for Ribosomal Protein Analysis
Service Overview
Ribosomal proteins (RPs) are not merely structural components of ribosomes, they orchestrate mRNA translation, regulate cell growth, and respond to stress signals. Our WB RPs analysis service provides a high-resolution platform to quantify RPs expression, modifications, and interactions across diverse biological contexts. By integrating optimized lysis protocols, antibody specificity validation, and advanced imaging systems, we enable researchers to:
- Detect low-abundance RPs with femtogram sensitivity.
- Resolve phosphorylated isoforms using post-translational modifications (PTMs) specific antibodies.
- Compare RPs dynamics in subcellular compartments.
Fig.1 Single-cell western blotting for single-cell protein analysis.1
Simplified Procedures
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Sample Submission & Customized Consultation
We accept diverse sample types, including mammalian/plant cells, tissues, and collaborate closely with clients to align protocols with research goals. Clients provide sample details to streamline customization, laying the foundation for reproducible results. -
Protein Extraction & Stabilization
Samples undergo lysis in RNase-free buffers supplemented with protease/phosphatase inhibitors to prevent ribosome degradation and preserve PTMs. Protein concentration is quantified via BCA/Bradford assays, with equal loading confirmed by Coomassie staining of replicate gels.
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High-Resolution SDS-PAGE
Proteins are separated on Tris-glycine, chosen to resolve ribosomal subunits with sharp precision. Molecular weight markers are included for accurate band identification, while Coomassie-stained replicate gels validate protein integrity and loading consistency.
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Efficient Protein Transfer & Blocking
Semi-dry blotting effectively immobilizes proteins onto PVDF or nitrocellulose membranes, with the success of the transfer being validated through reversible staining. Membranes are blocked with 5% BSA or non-fat milk in TBST, minimizing non-specific binding and ensuring clean signals during antibody incubation.
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Antibody Incubation & Dual Detection Methods
Primary antibodies are incubated overnight at 4°C for high specificity, with dilutions optimized per target. Secondary antibodies conjugated with horseradish peroxidase (HRP) for chemiluminescent detection or labeled with fluorophores for fluorescent detection facilitate multiplex analysis.
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Quantitative Imaging & BioinformaticsBlots are imaged, capturing high-resolution data for precise band quantification. Optional bioinformatics integrates RPs' data with pathways or disease contexts, adding depth to findings.
Key Applications
- Disease Diagnostics & Biomarker Discovery
Detect disease-specific proteins or their PTMs, such as phosphorylated RPs in cancer or neuroinflammatory markers in neurological disorders, to enable early diagnosis, prognosis, and personalized treatment strategies.
- Gene Expression Validation
Confirm translational regulation of genes by correlating mRNA levels with protein abundance, resolving discrepancies due to translational repression or PTM-mediated stabilization. Ideal for functional genomics studies.
- Signal Transduction Pathway Mapping
Unravel cellular signaling cascades by profiling phosphorylation states of key nodes, such as RPS6 in PI3K/AKT/mTOR signaling or subcellular translocation of signaling proteins, providing mechanistic insights into disease pathogenesis.
- Protein-Protein Interaction Networks
Identify RPs interactors or functional complexes via co-immunoprecipitation (co-IP) coupled with WB, revealing dynamic associations that regulate translation, apoptosis, or viral replication, such as SARS-CoV-2 NSP1 binding to RPL19.
- Subcellular Localization Studies
Resolve spatial distribution of RPs across compartments, such as cytoplasmic vs. nuclear fractions, using subcellular fractionation followed by WB, linking localization to functional roles in stress response or oncogenesis.
Distinctive Strengths
- Unmatched Expertise and Experience
With decades of experience, expert teams possess deep insights into troubleshooting common WB challenges， such as poor protein extraction, inefficient transfer, non-specific binding, high background, and delivering robust, reliable data. Their knowledge base extends to complex samples and specific protein types like phosphoproteins and membrane proteins, leveraging rigorous control of experimental workflows and precise electrophoretic separation strategies.
- Reliability and Reproducibility
Adherence to standardized and internally developed protocols, rigorous quality control measures at every step, and skilled execution ensure consistent and highly reproducible results, critical for publishable research and regulatory submissions.
- Cost and Time Efficiency
By leveraging specialized facilities, cutting-edge equipment, and experienced personnel, researchers can avoid the significant upfront investment in laboratory setup and the extensive time required for assay development and optimization. This allows in-house teams to focus on data interpretation, experimental design, and other core research activities.
- Access to Advanced Technology
Professional services often utilize state-of-the-art imaging systems and proprietary reagents to achieve superior sensitivity and lower background, leading to clearer, more quantifiable results. This includes advanced multi-channel fluorescence detection for multi-target analysis.
FAQs
Q: What does the WB RPs analysis service include?
A: Our service provides comprehensive WB analysis of RPs, including detection of expression levels, phosphorylation states, and subunit composition. We offer quantitative data validation and customizable report generation.
Q: How are results delivered?
A: Clients receive digital reports containing:
- Full-resolution blot images
- Densitometry quantification graphs
- Statistical analysis (p-values, fold changes)
- Biological interpretation of RPs expression patterns.
Q: Do you offer antibody validation?
A: Yes, we maintain a validated antibody panel targeting major RPs. Customized antibody testing services can be provided based on specific requests.
Q: How is data confidentiality maintained?
A: All client data is handled under strict NDA agreements. Raw images and processed data are securely archived for 6 months and permanently deleted upon request.
Q: Do you provide post-analysis support?
A: Yes, our team offers free consultation for result interpretation and can recommend complementary assays, such as polysome profiling, RNA-seq integration for follow-up studies.
WB analysis remains an indispensable technique for advancing our understanding of protein function in health and disease. For researchers and companies seeking accurate, reliable, and high-throughput protein detection and quantification, partnering with expert WB analysis services is a strategic decision. By entrusting your protein analysis needs to seasoned professionals, you can overcome technical hurdles, ensure data integrity, accelerate your discovery pipeline, and ultimately, unveil the intricate secrets held within cellular proteins. Connect with a trusted provider today to empower your research with precision protein analysis.
Reference
- Liu, Lixing, et al. "Advances of single-cell protein analysis." Cells 9.5 (2020): 1271. DOI: 10.3390/cells9051271. Distributed under Open Access license CC BY 4.0 , without modification.
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