Services, instrumentation and equipment

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Mass spectrometry services

We offer mass spectrometry services such as:

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)

  • LC/MS analysis is performed using a Thermo Scientific Orbitrap Fusion with ETD coupled to a Thermo UltiMate 3000 nanoLC system and a New Objectives digital PicoView source.
  • Protein Identification from excised 2D PAGE or DIGE gel plugs is performed using a quick LC gradient of 30 minutes.
  • Protein Identification from excised 1D PAGE gel plugs is performed using an LC gradient of 60- 90 minutes.
  • Proteome characterization is performed using Tandem Mass Tags and is analyzed using multi-dimensional HPLC. The first dimension is basic RPHPLC followed by traditional reversed phase HPLC. The first dimension is fractionated using a Thermo Scientific UltiMate 3000 LC system. Collected fractions are then analyzed using the Fusion over a two hour gradient.
  • Targeted proteomics are also performed using the Thermo Scientific Fusion mass spectrometer.
  • Data analysis is performed using Thermo Scientific’s SEQUEST® and Proteome Discoverer. Data output is provided to the user as a Scaffold output file and Ingenuity Pathway Analysis.

Requirements

>50 fmol of tryptic digest on column; sample in an aqueous solution of <5% acetonitrile, <0.01% (w/v) detergent concentration.

Mass spectrometry instrumentation and sample prep equipment

The Thermo Scientific Fusion and other coupled equipment

Thermo Scientific Fusion

Equipped with a heated New Objectives DPV-565 Digital nanospray source allowing for the use of extra long columns up 50 cm. Coupled to a Thermo Scientific UltiMate 3000 NanoLC

The Thermo Scientific Vantage and other coupled equipment

Thermo Scientific Vantage

Equipped with a Thermo Flex Flex Nanospray source. Coupled to a Thermo Scientific UltiMate 3000 LC.

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.