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Proteomics – Identification and Relative Quantification of Proteins

In the field of proteomics, our team supports identification and quantification of proteins. Feel free to contact us if you want to know the composition of proteins in a sample or if you are interested to compare differences between various groups of samples. We are happy to find the appropriate analysis method for your specific research question together with you, no matter what you are working with, be it complex cell lysates, immunoprecipitated samples or protein analysis using 1- or 2-dimensional SDS-PAGE gels. Proteomics services are provided in “Full service mode” or “Partial service mode” if protein digest and/or data evaluation are performed by experienced users themselves.

 

Our service

Service fees comprise material and equipment running costs starting from submission of the protein lysate or 1D gel band/2D spots to the report. In detail our service includes:

  • Help and advice during project planning and application for grants
  • Sample preparation for mass spectrometric analysis: Digest of extracted proteins with various digestion protocols (FASP, SP3, S-Trap, in-gel, in-solution)
  • Analysis of peptides using nano-HPLC coupled to high-resolution mass spectrometers (Bruker timsTOF HT or Thermo Orbitrap QExactive HF)
  • Relative label free quantification
  • Database search in available protein databases (from public servers as NCBI or UniProt) as well as your own protein database in fasta format
  • A detailed report of your results containing all the information you need for publication. Transfer of raw data files to be uploaded to public repositories by the users
  • We also provide basic statistical analysis (T-test, ANOVA) if required. For any further statistical analysis, you are kindly advised to contact a biostatistician
  • Help and advice during manuscript writing

Available instruments

The timsTOF HT (Bruker Daltonics) is a quadrupole time-of-flight mass spectrometer with integrated trapped ion mobility separation (tims). It is coupled to an Evosep One HPLC system (Evosep Aps) via a CaptiveSpray 2 nano-ESI source for standardised peptide separation. Equipped with tims and Parallel Accumulation Serial Fragmentation (PASEF), this instrument setup enables high-throughput mass spectrometry for large-scale qualitative and quantitative proteomics analyses supporting our collaboration partners. The system is offered in full- or partial service mode and enables:

  • High-throughput proteomics analyses
  • Protein identification (Shot gun proteomics)
  • Quantitative proteomics (Label free quantification)
  • Relative protein quantification
  • Data-independent acquisition

Ion mobility as a fourth dimension of separation

The Q Exactive HF Orbitrap system (Thermo Fisher Scientific) is a high-resolution mass spectrometer coupled to an Ultimate 3000 nanoRSLC system (Dionex) via a nano-electrospray ionization source (nESI). The system combination is used for proteomics analyses (protein identification and quantification) performing data-dependent acquisition in full- or partial-service mode. The system enables:

  • Proteomics analyses
  • Protein identification (Shot gun proteomics)
  • Quantitative proteomics (Label free quantification)
  • Relative protein quantification
  • Data-dependent acquisition