EQUIPMENT

In 2004 the Proteomics Core and the Duncan Lab relocated to the Anschutz Medical Campus where they now jointly occupy approx. 2,500 sq. ft. of purpose-designed space on the 1st and 12th floors of Research Complex 1 (south). Together, the Proteomics Core and Dr. Duncan's laboratory include the following instruments:

Eksigent nanoHPLC autosampler
This autosampler increases the sample analysis capacity and utilization of an ESI MS/MS instrument by as much as 300%. The autosampler has the capacity to deliver 0.1 µL - 1 mL per sample, in 0.1 µl increments, from up to two micro-plates (96 or 384 wells). Direct transfer of micro-plates from the digestion robot to the autosampler for MS data acquisition further minimizes sample handling and improves throughput.

GE Healthcare Ettan MDLC
This is a multidimensional chromatography system that optimizes micropreparative protein and peptide prefractionation as well as nanoscale peptide separation. The MDLC provides accurate and reproducible flow rates and gradients, with flow rates ranging from 200 nl/min to 2 ml/min.

Applied Biosystems Voyager STR+ MALDI-TOF
A matrix-assisted laser desorption ionization time-of-flight mass spectrometer with delayed extraction for improved ion focusing and post source decay (PSD) capabilities for peptide sequencing.

Applied Biosystems Voyager DE-PRO MALDI-TOF
A bench-top version of the STR+. A matrix-assisted laser desorption ionization time-of-flight mass spectrometer with delayed extraction and post source decay capabilities. A high voltage detector is employed in the linear mode; a second detector is employed in the reflectron mode.

Finnigan LCQ-Deca
This instrument is fitted with a custom-built nanospray source for capillary LC-nanospray MS operation and has an Eksigent LC pump with autosampler. The LCQ is capable of data dependent LC-MS/MS analysis and is well suited to proteomics work.

Applied Biosystems Q-STAR XL
This is a tandem quadrupole time-of-flight (Q-TOF) configuration instrument ideally suited to high sensitivity peptide characterization. The accurate mass capability of this MS/MS instrument allows for de novo sequencing of peptides, and with on-line HPLC separation, this system offers accurate, rapid and sensitive peptide and protein identification. This system is fitted with a nanospray source and an Eldex MicroPro HPLC system.

HP 5973 GC-MSD
A fully automated bench-top gas chromatograph-mass spectrometer used for the quantification of posttranslationally modified amino acids (e.g., oxidized phenylalanine) in biological samples.

Agilent LC/MSD Trap SL
An ion trap mass spectrometer with multiple ion sources available including nanospray, electrospray and atmospheric pressure MALDI. This system is capable of data dependent LC-MS/MS analysis and is well suited to proteomics work.

2-D Gel Electrophoresis Equipment
The Duncan lab has seven separate electrophoretic assemblies (3 GE Healthcare IPGPhor isolectric focusing units and 4 second dimension units, an GE Healthcare Ettan Dalt Twelve and 3 BioRad Protean xi systems).

GE Healthcare Typhoon 9400 The Typhoon is a variable mode laser scanning imager for fluorescence imaging with a broad dynamic range and has a low limit of fluorescence detection. This system allows us to make full use of Ettan DIGE, 2-D fluorescence difference gel electrophoresis (DIGE)

GE Healthcare DeCyder Differential Analysis Software
This software suite includes the Difference In-gel Analysis (DIA), Biological Variation Analysis (BVA) and Extended Data Analysis (EDA) packages. Together, these programs provide spot detection, relative quantification of normalized spot volume and spot comparison across multiple gels.

GE Healthcare Ettan Spot Picker
The Ettan Spot Picker is a robotic system designed to accurately pick protein spots from 2-D gels and transfer the plugs to microplate wells. This unit works in concert with the Ettan Digester and has the capacity to pick 9600 protein spots per batch run.

GE Healthcare Ettan Digester
The Ettan Digester performs in-gel digestion of plugs collected from 2-D gels. The Digester automatically tracks samples from spot picking to completed digestion in bar-coded 96-well plates.

Randox Evidence Biochip Array System
The Evidence system is a fully automated, continuous access immunoanalyser using protein biochip array technology. Immunoassays for multiple analytes are performed simultaneously on the biochip surface. Assay endpoints are chemiluminescent reactions captured and quantified by a CCD camera for excellent intra- and interassay precision. It is currently the only FDA approved protein biochip instrument.