Services, instrumentation and equipment

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Mass spectrometry services

We offer mass spectrometry services such as:

  • Gel, IP and solution based protein identification
  • Proteome quantitation using both isobaric labelling (TMT) and label free methodologies
  • Post-translational modification analysis
  • Sample preparation

Enzymatic protein digestion

Before being analyzed by mass spec, most proteins must be broken into smaller fragments. This is accomplished by digestion with a protease, usually trypsin and Lys-C, but may be another proteases of your choice.

Requirements

>100 fmol protein, may be a protein in solution, a gel slice or spot that is visible by coomassie staining.

Liquid chromatography/mass spectrometry (LC/MS)

  • Thermo Scientific Orbitrap Eclipse with ETD, coupled to a Thermo Ultimate 3000 nanoLC system.
  • Bruker timsTOF Pro2, coupled to a Bruker NanoElute LC.
  • Evosep One LC is primarily used for clinical samples such as whole blood, serum, and urine. Can be coupled with either the Eclipse or TimsTOF depending on experiment
  • Protein Identification can be performed on either gel bands or solution.
  • Proteome characterization is performed using either label free or isobarically labeled approaches.
    • Tandem Mass Tags (TMT) are analyzed using multi-dimensional HPLC. The first dimension is basic RPHPLC followed by traditional reversed phase HPLC on the Eclipse.
    • Data-Independent Acquisition (DIA) is performed using either the Eclipse or the timsTOF and the data processed using Biognosy's Spectronaut software.
  • Targeted proteomics is performed on either the Eclipse or timsTOF.
  • Data analysis is performed using either Biognosys Spectronaut or Thermo Scientific’s SEQUEST® and Proteome Discoverer. Data output is provided to the user through either Spectronaut or Proteome software Scaffold with both having free viewers.

Requirements

>50 fmol of tryptic digest on column; sample in an aqueous solution of <5% acetonitrile, <0.01% (w/v) detergent concentration. Contact the Center to discuss needs or questions.

Mass spectrometry instrumentation and sample prep equipment

Visit Nevada INBRE for information about grant citations. In addition, samples analyzed using either the Bruker timsTOF Pro2 or the Evosep One LC should also acknowledge the National Institutes of Health Shared Instrumentation Program (1S10OD028530-01A1).

Thermo Scientific UltiMate 3000 LC with integrated fraction collector

Thermo Scientific UltiMate 3000 LC

With integrated fraction collector.

nanoLCMS High Pressure Packing Pump

nanoLCMS High Pressure Packing Pump

Custom packing of columns with Dr. Maisch Reprosil-Pur 1.9um packing up to 50cm in length for maximum performance.