PROTEIN ANALYSIS

LJIDI has shared equipment for routine Western Blotting, magnetic immunoprecipitations, and gel staining for protein (Commassie, silver, and Sypro).  A gel imager (UVP AutoChem) measures chemi-luminescence from Western Blots as well as images gels stained with fluorescent dyes, such as Sypro.  A Beckman P/ACE with fluorescence detection is available for the reproducible analysis of proteins and amino acids.  A Fisher Sonic dismembrator disrupts cells and tissue for protein analysis.  A plate reader (BioTek Epoch Multi-Mode) is available for ELISA, fluorescence, time-resolved fluorescence and luminescence meausrements. We have migrated most of our ELISA protein analysis assays onto the microsphere/flow cytometry platform (Accuri) because microsphere-based assays are sensitive, and are readily multiplexed to improve throughput. 

A Nanophotometer P360 (Implen) measures protein and nucleic acid concentrations, hemoglobin levels and state, and cell numbers in volumes ranging between 0.3 and 4 µl. 

The CFAR partially sponsored Protein Expression and Proteomics Core (TSRI, Dr. Bruce Torbett, Co-Director) has available proteomics equipment and instrumentation. In the core available are: A Bio-Plex instrument, 1- and 2-D gel electrophoresis protein separation instrumentation, IEF Zoom fractionator for solution phase IEF, state of the art image 2D Proteomic Imaging System with a Bio-Rad VersaDoc 1000 and Typhoon 3-color laser imagers, and Progenesis SameSpots and Quantity One/PDQuest software with additional websites, and a ProPic II automated spot picker, and ProGest protein digestion unit. The CFAR core has available a Thermo LCQ Deca XP MAX LC/MS/Msn ion trap instrument, with an automated sampler, to analyze protein samples obtained from 1- and 2-D GE or protein analyses in MudPIT (2D-nanospray) mode. Assorted protein analysis software, such as SEQUEST, DTASelect, PROTOMAP, GutenTag, SALSA, and DaughterDB, and CPU time is available. the core has access to a Thermo LTQ Orbitrap, and other instruments, for increased mass accuracy, mass resolution and identification of proteins / compounds in complex mixtures (Center for Physiological Proteomics, Dr. John Yates).