Selected reaction monitoring (SRM), also known as multiple reaction monitoring (MRM), is an important technique for quantitative of target proteins in complex sample. SRM/MRM technique can achieve rapid, sensitive, specific quantification of the target protein in a complex system and excellent quantitative reproducibility. SRM/MRM technology eliminates most of the non-target detection, that making the noise signal greatly reduced, and improves the detection sensitivity significantly. Compared to ELISA, SRM/MRM has better reproducibility, higher throughput, lower development cost and higher stability. At present, SRM/MRM has rapidly evolved into the representative technology of target technology, and is regarded as the "gold standard" for protein quantification based on mass spectrometry.
SRM/MRM technique is based on the known or assumed reactive ion information, targeted to select the data for mass spectrometry signal acquisition, removal of non-compliance with the interference of ion signals, through the data Mass spectrometry to obtain mass spectrometric quantitative information. The main instrument used in this technique is a triple quadrupole mass spectrometer, in which the first and third quadrupole are used as mass filters to specifically select peptide ions (precursor ions) that are identical to the pre-set mass to charge ratio. And the fragment ions (product ions) through the second quadrupole as the collision cell by CID (Collision Induced Dissociation) way to break the precursor ion to produce ions. SRM technology has a high selectivity, reduced the background noise, and interference of ions.
Highlight of SRM/MRM technique:
- High reproducibility;
- High throughput;
- No antibodies;
- Gold standard for targeting quantitative.
SRM/MRM Analysis Services at Creative Proteomics offers you a state-of-the-art strategy of proteins analysis. The workflow is based on the following sections:
- The SRM/MRM assay development. This is the core of the approach. First, 2 or 3 targeted peptides should be selected from a spectral library generation or Skyline software. The selected peptides should be unique to the protein of interest and easily detected by LC-MS. They also require no missed cleavage sites and no frequently modified amino acids. The product ions should also be selected. Second, detection conditions, such as fragmentation energy and cycle time, should be optimized. Third, a working linear curve was built with the corresponding concentration ratios on the x-axis and peak area ratios on the y-axis.
- Analysis of the sample. After digestion, the sample is performed on the triple quadrupole mass spectrometer.
- Data analysis. Data will be analyzed with Skyline software.
SRM/MRM application
- Verification of iTRAQ differential protein;
- Verification of label-free differential protein follow-up;
- Protein and peptide absolute quantification;
- Quantification of disease markers, the establishment of diagnostic models;
- Phosphorylation protein quantification, methylation protein quantification;
- Other post-translationally modified protein quantification;
- Quantitative analysis of pathways.
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As one of the leading omics industry company in the world! Creative Proteomics now is opening to provide proteomic analysis of post-translational modifications service for our customers. With over 8 years of experience in the field of bioinformatics, we are willing to provide our customer the most outstanding service! Contact us for all the detailed information!