ANALYTICAL PROTEIN PROFILING

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This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Proteomics Lab  Michael Powell, Leader The main instrumentation of the Proteomics core lab consists of: 1. A ThermoFinnigan LTQ mass spectrometer coupled to an Agilent 1100 HPLC system to provide LC/MS/MS analysis. 2. A BioRad automated spot cutter (Proteome Works) to allow the differential analysis of 1D and 2D gels. 3. A BioRad FX Imager that allows the imaging of 2D DIGE gels and other fluorescently labeled gels. 4. A BioRad BioPlex 100 bead sorter, which allows the performance of multiple analyte assays using Luminex bead technology. Analytical Chemistry Lab  Daniel von Deutsch, Leader The newly renovated MSM ACL is equipped with multiple means to perform chromatographic and spectrographic analysis of biological samples. High Performance Liquid Chromatography (HPLC). The ACL has two Agilent (HP) 1100 series HPLCs. The available detectors include: 1) Diode Array Detector (DAD), 2) Variable Wavelength Detector (VWD), 3) 1046A Fluorescence Detector (FD), 4) an 1100 series Fluorescence Detector (FD). Optional detectors include a high sensitivity Gilson fluorescence detector, an Agilent 1049A Electrochemical Detector (ECD) and an ESA Coulochem II ECD system outfitted with multiple Model 5011 high sensitivity two channel analytical cells. Gas Chromatography/Mass Spectrometry (GC/MS). In addition, the ACL is equipped with two Agilent (HP) GCs for the detection of drugs and their metabolites. These include an Agilent (HP) 6890 Series capillary GC with Flame Ionization Detector (FID) and Nitrogen Phosphorus Detector (NPD) and an Agilent (HP) 5890 Series capillary GC coupled with 5972 Series Mass Spectrometer (MS - EI). Data can be acquired in the SCAN or Selected Ion Monitoring (SIM) modes. Spectrophotometry. The ACL is equipped with a Spectromax Plus is a UV/VIS variable wavelength microtiter plate reader with Softmax Pro software used for performing colorimetric spectrophotometry and immunoassays. This equipment allows the ACL to offer qualitative and quantitative analysis of small molecules from either human or animal samples. In addition to standard pharmacological or biochemical testing, the ACL is also capable of performing other tests that includes: drug identification, chiral separations and analysis, and redox analysis (determining levels/ratios of reduced and oxidized molecules such as glutathione). Service Charges: Proteomics Lab/2D Gel Lab We currently have a fee schedule for all services. For example, we charge by the LC/MS/MS run for analysis on the LTQ. This charge includes a digestion of the sample by trypsin, an LC/MS run, and the subsequent database searches. Database searches including post translational modifications incur an additional charge for typical modifications such as phosphorylation, nitration, ubiquitination, etc. Other more complicated analysis are charged from an estimate of the time required. Luminex assays are offered from any manufacturer. The charges are based on the price of the kit plus a setup fee of $200 per assay. Analytical Chemistry Lab The Analytical Chemistry Laboratory has a standard fee schedule for services. The fee structure is based on the amount of time required to process the samples (extraction, derivatization, etc) and the cost of supplies and reagents. In addition, a general fee is requested to help cover equipment maintenance and costs of operation.