Label-dependent Strategies for Ribosomal Proteome Identification Service

Creative Biolabs is a leading service provider that focuses on ribosomal proteomics research. Based on our advanced platforms and extensive experience, now we can provide a series of label-dependent strategies for ribosomal proteome identification.

Background

The research of ribosomal proteomics consists of four basic procedures, including ribosome isolation, ribosomal proteome separation, ribosomal proteome identification, and candidate protein validation. In each link, various strategies have been developed to improve the reliability and efficiency of research.

The radical-free and highly reducing (RFHR) 2-DE method is an optimal alternative for separating ribosomal proteins (RPs) that has been successfully performed in several ribosomal proteomic studies. However, ribosomal proteome identification performed only by RFHR 2-DE presents various disadvantages, such as accuracy and scale limitations. In this case, Creative Biolabs describes a range of MS-based label-dependent strategies for ribosomal proteome identification.

Fig. 1 Overview of clinical cancer proteomics strategies.¹

Our Service

Creative Biolabs offers specialized label-dependent strategies for ribosomal proteome identification service, leveraging our advanced platforms and deep expertise in ribosomal proteomics research. Our label-dependent approaches, including cutting-edge MS-based methods, deliver reliable and efficient solutions for ribosomal proteome identification. With advancements in mass spectrometry (MS) technology, label-dependent strategies such as ICAT and SILAC have become fundamental in ribosomal proteomics.

1. ICAT (Isotope-Coded Affinity Tag)

During ICAT quantification, proteins extracted from two different cell types are labeled with chemical tags that possess varying isotopic compositions yet maintain the same structural properties. Following hydrolysis, the labeled peptides are purified using Avidin and then analyzed via mass spectrometry. Tandem mass spectrometry is employed to identify the peptide sequences and the corresponding types of proteins. In the primary mass spectrometer, a signal pair with a fixed mass difference is observed for each peptide tagged with light and heavy isotopes. This signal pair enables the determination of the relative expression levels of the same protein across different cellular states.

2. SILAC (Stable Isotope Labeling by Amino Acids in Cell Culture)

SILAC technology employs a culture medium enriched with light, medium, or heavy isotope-labeled essential amino acids, primarily lysine and arginine, to culture multiple groups of cells and label the newly synthesized proteins within them. Typically, after 5-6 generations of culture, all proteins in the cells become fully isotope-labeled, effectively replacing the original amino acids. This approach results in a difference in molecular weight between the two sets of proteins, while all other chemical properties remain unchanged. The two groups of cells are then mixed, and following protein collection and trypsin digestion, the SILAC-labeled samples are analyzed using LC-MS/MS. By comparing the peak areas of the labeled peptides, researchers can accurately quantify protein abundance.

Service Process

Sample Requirements

• Please ship frozen cells using dry ice, ensuring that the cells can be subcultured more than 5 times.
• Ship the labeled and prepared cell samples or protein samples on dry ice, with each sample containing 50-100 µg of protein.

Highlights

• Comprehensive Label-dependent Approaches

We offer a variety of label-dependent strategies, including SILAC and ICAT, to enhance the precision and reliability of ribosomal proteome identification.

• Advanced Mass Spectrometry (MS) Integration

Our service leverages the latest MS technology, ensuring high sensitivity and accuracy in ribosomal proteomics research, even for complex samples.

• Enhanced Protein Quantification Accuracy

By utilizing stable isotope labeling, our approaches enable precise quantification of ribosomal proteins, allowing for detailed comparative studies.

• Tailored for Ribosomal Proteomics

Our methods are specifically adapted for the unique needs of ribosomal proteome analysis, overcoming common challenges such as the variability in ribosomal protein composition.

• Extensive Ribosomal Research Expertise

Backed by years of experience in ribosome research, our team provides expert guidance and solutions, ensuring reliable results tailored to your specific research needs.

• High-throughput and Scalable Options

Designed for scalability, our service can accommodate high-throughput projects, making it ideal for large-scale ribosomal proteomics studies.

• Dedicated Technical Support

Our scientists offer ongoing support throughout the project, from initial planning to data analysis, ensuring seamless execution and valuable insights into ribosomal proteomics.

FAQs

Resource

Ribosome Analysis - Creative Biolabs Ribosome Analysis - Creative Biolabs

Creative Biolabs has been a long-term expert in the field of ribosome research. Based on our extensive experience and advanced platforms, we are confident in offering a series of ribosome-related services, including ribosome separation and extraction services, ribosome analysis services, as well as ribosomal marker antibody development services. If you are interested in our products or services, please do not hesitate to contact us for more detailed information.

Reference

1. Macklin, Andrew, Shahbaz Khan, and Thomas Kislinger. "Recent advances in mass spectrometry based clinical proteomics: applications to cancer research." Clinical proteomics 17.1 (2020): 17. Distributed under the Open Access license CC BY 4.0, without modification.