MALDI-TOF Mass Spectrometry (Link)
The MALDI-TOF facility housed in the Department of Biochemistry provides access to mass spectrometric fingerprinting of unknown proteins. MALDI-TOF (Matrix-assisted, Laser-Desorption-Ionization/Time of flight) mass spectrometry is presently the method of choice for identification of unknown proteins via mass analysis of proteolytic peptides, and for characterization of post-translational modifications. This technique is rapid, highly sensitive, and applicable to a wide variety of research problems. Applications include direct characterization of mutated proteins, estimating the extent of protein derivatization (e.g., biotinylation), and identification of unknown proteins isolated from polyacrylamide gels. Depending on the specific application and complexity of the system, reliable data can be obtained in the fmol-pmol range.
2D-DIGE (Differential In-Gel Electrophoresis) allows extracts of treated and untreated cells to be co-electrophoresed on the same gel and then quantified, thereby avoiding problems associated with registering independently-derived gel maps. This technique depends on pre-derivatization of samples with a fluorescent Cy-dye (Cy-2, Cy-3, or Cy-5). These dyes are MW and charge-matched, have non-overlapping excitation and emission spectra, and can be covalently attached to amino groups of proteins via an NHS ester substitution reaction.
BIAcore X (Link)
The BIAcore facility housed in the Department of Biochemistry provides access to a highly sensitive method to measure and analyze macromolecular interactions. The facility currently contains a BIAcore X instrument that utilizes surface plasmon resonance to measure biomolecular interactions in real-time in a label-free environment. One of the interactants is immobilized to the sensor surface and the others are free in solution and passed over the surface. Association and dissociation kinetics are measured in arbitrary units and displayed in a graph called the sensorgram (see http://www.biacore.com/lifesciences/index.html for additional details).
