Isobaric Label

DESCRIPTION
SPECIFICATION
RESOURCES

Service Description

We have developed innovative solutions for optimizing the process of accurately quantifying proteins in complex biological matrices and other complex mixtures.

Isobaric Label TMT Quantitation Proteomics

Isobaric Label services incorporate sample multiplexing to provide deep proteome coverage and highly precise quantitation of small or medium-sized sample sets. Samples are prepared and labeled separately, then all samples are pooled and analyzed by nano-flow LC-MS/MS1.

Project Workflow

BGI Genomics provides Isobaric Label Quantitative Proteomics service using iTRAQ (Isobaric Tags for Relative and Absolute Quantitation), IBT (Isobaric Tags for Quantitation) and TMT (Tandem Mass Tags for Quantitation) technologies developed by AB SCIEX, BGI and Thermo Scientific, respectively.
  • Protein extraction
  • Protein digestion
  • Isobaric labeling
  • HPLC fractionation
  • LC-MS/MS (DDA)
  • Data analysis

Reference

[1] Wong, J. W. H., & Cagney, G. (2009). An Overview of Label-Free Quantitation Methods in Proteomics by Mass Spectrometry. Proteome Bioinformatics, 273–283.doi:10.1007/978-1-60761-444-9_18.

Mass Spectrometry Service Specification

Isobaric Label Quantitative Proteomics services are performed using nano-flow liquid chromatography and cutting-edge mass spectrometer (Q Exactive HF/HF-X).

  • Sample Preparation and Services

    • The QC of enzymatic hydrolysis and labeling steps are strictly controlled by LC-MS DDA analysis
    • 1h/fraction; 20 fractions for eukaryotes;10 fractions for prokaryotes

  • Quality Standard

    • Summary includes all methods and data analysis
    • Reports provided in Excel or PDF format, RAW files available upon request

  • Turn Around Time

    • Typical 4-5 weeks from sample QC acceptance to data report delivery

Sample Requirements

SAMPLE TYPE AMOUNT AND CONCENTRATION MINIMUM SAMPLE VOLUME
Protein sample in liquid solution Recommended 1000 µg: 2 µg/µL 250 µL
Protein sample in liquid solution Minimum Required 50 µg: 1 µg/µL 50 µL

Data Analysis

DATA ANALYSIS
  • Data analysis and validation performed with Mascot
  • Protein identification and quantification list
  • Differential proteins data statistics and volcano plot
  • Principal component analysis (PCA) and cluster analysis
  • Protein GO/COG/KOG/Pathway annotation
  • GO/COG/KOG enrichment analysis of differential proteins
  • Protein-protein interaction analysis
  • Protein subcellular localization analysis
  • Repeatability analysis

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