Home » Research

Proteomics & Molecular Characterization Core Facility

The Proteomics & Molecular Characterization Core Facility is a state-of-the-art laboratory designed to aid researchers in the separation, quantitation, identification, and characterization of expressed proteins in any given biological sample. Using the techniques available in this laboratory, a researcher will be able to ascertain what proteins are being expressed in any particular sample type under any given conditions, to what extent each of these proteins is being expressed, and finally, to determine the presence of any post-translational modifications in proteins of interest. The laboratory is equipped with the following instrumentation to allow such analyses.

Protein Separation:

Proteins can be separated either by 2-dimensional polyacrylamide gel electrophoresis (2D-PAGE) or by 2-dimensional chromatography (2D-LC). The first dimension of the 2D-PAGE is performed on an IPGphor, followed by separation in the second dimension on a DALT 6 gel electrophoresis apparatus. The gels are subsequently imaged on a Typhoon 9400 fluorescent imager (all from Amersham Biosciences). 2-dimensional chromatography is performed on a Surveyor multidimensional chromatography system (ThermoElectron).

Protein Quantitation:
For samples separated using 2D-PAGE, comparative protein expression analysis between multiple samples is performed using Differential In-Gel Electrophoresis (DIGE) followed by analysis with DeCyder 5.0 software (Amersham Biosciences).

2D-PAGE Protein Spot Processing:
Upon analysis of the spot pattern of single or multiple gels within an experiment, protein spots of interest are prepared for mass spectral analysis by the Spot Handling Workstation (Amersham Biosciences). This medium-throughput fully-automated robotic platform prepares up to 1152 protein spots for mass spectral analysis at a time.

Protein Identification:
The Proteomics & Molecular Characterization Core Facility has both a MALDI-ToF/Pro (Amersham Biosciences) and a ProteomeX Electrospray Ion-Trap (ThermoElectron) mass spectrometer for identification of proteins of interest using either Peptide Mass Fingerprinting techniques or direct peptide sequence information.

Protein Characterization:
Using either the MALDI-ToF/Pro or the ProteomeX mass spectrometer, primary sequence information can be determined for proteins and peptides of interest. Further, the presence or absence of specific post-translational modifications (PTM’s) can be probed.

Contact: proteomics@mail.jci.tju.edu