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Label-Independent Strategies

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-independent strategies for ribosomal proteome identification.

Strategies for Ribosomal Proteome Identification

With the continuous improvement of mass spectrometry (MS) technology, a series of label-dependent strategies have been developed rapidly for ribosomal proteomics research. For example, multiple reaction monitoring (MRM) is the most frequently used method. Compared with the label-dependent MS-based strategies, label-independent strategies have no advantages in terms of sensitivity and reproducibility. But at the same time, label-independent strategies are less likely to interfere with MS identification because it is not labeled.

In ribosomal proteomics research, by using linearly correlated MS information of different concentrations of peptides, such as m/z, spectral counts, or peak measurements, the nature and relative quantities of specific peptides can be determined. In recent years, we have successfully combined RFHR 2-DE with LC-MS/MS for ribosomal proteome analysis. In addition, multiple reaction monitoring (MRM) is also a tag-independent MS-based protein identification strategy, mainly for small-scale validation of interesting candidate proteins.

Fig. 1 Untargeted MS analysis to identify the number of peptides and proteins in 20 mouse organs and tissues. (Michaud, Sarah A., et al., 2024)Fig. 1 Number of peptides and proteins identified by untargeted MS analysis of 20 mouse organs and tissues.¹

MRM for Candidate Protein Validation

After identifying interesting candidate proteins, these proteins need to be validated to determine that their differential expression is objective and that these proteins truly belong to the ribosomal proteome. Western blotting and enzyme-linked immunosorbent assay (ELISA) are traditional methods for candidate protein validation. However, the necessity of specific commercial antibodies largely limits their use. In this case, MRM is an antibody-free method to identify predefined peptides derived from target proteins by filtering large numbers of interfering ions using certain tandem MS instruments. What's more, linking a large number of MRM transitions into a single assay enables the simultaneous quantification of hundreds of peptides and proteins.

Advantages of Label-independent Strategies

  • Interference with MS identification is avoided
  • Well-trained scientists and extensive experience
  • Low price and high-quality service
  • Rapid turnaround time

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. Michaud, Sarah A., et al. "Multiple reaction monitoring assays for large-scale quantitation of proteins from 20 mouse organs and tissues." Communications Biology 7.1 (2024): 6.
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