iTRAQ-Based Peptidomics Analysis Service

Creative Proteomics is a leading provider of biological quantitative analysis services for peptide analysis. Creative Proteomics utilizes state-of-the-art instrumentation to efficiently and accurately evaluate peptides. Our liquid separation laboratories are equipped with ultra-high performance liquid chromatography (UPLC), high-performance liquid chromatography (HPLC), and liquid chromatography-mass spectrometry (LC-MS) instruments for the separation, identification, and quantification of peptides.

What is iTRAQ-Based Peptidomics Analysis?

iTRAQ-Based Peptidomics Analysis is a mass spectrometry-based approach used for quantitative analysis of peptides in biological samples. iTRAQ (isobaric tags for relative and absolute quantitation) is a chemical labeling technique that allows for simultaneous multiplexed quantification of peptides from different samples. In iTRAQ-based peptidomics analysis, the peptides are first extracted from the sample and then labeled with iTRAQ reagents. Each iTRAQ reagent consists of a specific mass tag that is attached to the peptide, allowing for distinction and quantification of peptides from different samples during mass spectrometry analysis. The quantitative information is obtained by measuring the intensity of the iTRAQ reporter ions generated during fragmentation of the labeled peptides. The abundance levels of the peptides in different samples can be compared by comparing the relative intensities of the reporter ions. iTRAQ-based peptidomics analysis is widely used in proteomics and peptidomics research to study changes in peptide expression levels and identify potential biomarkers in various biological samples.

iTRAQ-Based Peptidomics Analysis Services by Creative Proteomics

iTRAQ (Isobaric Tags for Relative and Absolute Quantification) is a popular technique used for quantitative analysis of peptides in proteomics and peptidomics research. It allows for simultaneous identification and relative quantification of multiple samples in a single experiment.

• Sample Preparation
  It starts with the extraction of peptides from the biological sample, which can be tissues, cells, or biofluids. The extracted peptides are then subjected to enzymatic digestion to produce smaller peptide fragments.
• iTRAQ Labeling
  The peptide fragments are labeled with iTRAQ reagents. These reagents contain unique isobaric tags that enable multiplexing of samples. Each iTRAQ label has a specific mass that allows for relative quantification of peptides from different samples.
• Peptide Fractionation
  To reduce sample complexity, peptide fractions are often generated using various techniques such as high-performance liquid chromatography (HPLC).
• Mass Spectrometry Analysis
  The labeled peptide fractions are analyzed using tandem mass spectrometry (MS/MS). This involves ionizing and fragmenting the peptides to obtain their mass-to-charge ratios (m/z) and sequence information. The iTRAQ tags also provide quantitative data on the relative abundance of peptides across different samples.
• Interpretation and Reporting of Results
  Draw conclusions based on analysis results and generate relevant reports.

Techniques and Instrumentation for iTRAQ-Based Peptidomics Analysis Services

• Peptide Fractionation
  This technique involves separating complex peptide mixtures into several fractions using liquid chromatography techniques such as high-performance liquid chromatography (HPLC). It helps in reducing sample complexity and enhancing the detection of low-abundance peptides.
• Liquid Chromatography-Mass Spectrometry (LC-MS): It combines liquid chromatography with mass spectrometry to separate, ionize, and detect peptides based on their mass-to-charge ratio (m/z).
• Thermo Scientific Q Exactive Series
• SCIEX TripleTOF Series
• Agilent 6550 iFunnel Quadrupole Time-of-Flight (Q-TOF)
• Waters SYNAPT G2-Si HDMS System
• Tandem Mass Spectrometry (MS/MS): It involves fragmentation of peptides in the mass spectrometer, which provides sequence information for peptide identification.
• AB SCIEX TripleTOF 5600/6600
• Thermo Scientific Orbitrap Fusion/Lumos
• Agilent 6490/6550 Triple Quadrupole
• Waters Xevo TQ-XS
• Bruker Impact II/maXis II/QTOF
• Shimadzu LCMS-8060/9030 Triple Quadrupole

Applications of iTRAQ-Based Peptidomics Analysis

• Identification and quantification of differentially expressed peptides in biological samples
• Comparative analysis of peptide abundance in different biological conditions or disease states
• Profiling and characterization of peptide modifications or post-translational modifications
• Investigation of protein-protein interactions and protein complexes
• Quantitative analysis of proteolytic processing events and protein maturation
• Discovery of novel peptides and biomarkers
• Assessment of peptide dynamics and turnover kinetics
• Study of peptide-protein interactions and binding affinities

Our Advantages

• Multiplexing Capability: iTRAQ allows for the simultaneous quantification of multiple samples, typically up to 8 or 10, using isobaric tagging.
• Relative Quantification: iTRAQ provides relative quantification of peptides, allowing researchers to compare the abundance of peptides across different samples.
• High Sensitivity: iTRAQ-based peptidomics analysis combines high-performance liquid chromatography (HPLC) with tandem mass spectrometry (MS/MS), resulting in high sensitivity and the ability to detect low-abundance peptides.
• Comprehensive Peptide Profiling: iTRAQ analysis captures a broad range of peptides, providing a comprehensive view of the peptidome.

* For research use only. Not for use in diagnostic procedures!

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