Core E will provide state-of-the-art protein analysis services for SCCOR projects as a high quality, centralized[unreadable] resource to provide consistency and reproducibility in sample preparation, data analysis, and cost savings by[unreadable] eliminating duplications in equipment and reagents. Core E is based in the Biomolecular Resource Facility (BRF),[unreadable] an established Institutional Core Laboratory of the University of Texas Medical Branch that is partially funded by[unreadable] the NHLBI Proteomics Center contract mechanism. The BRF occupies 6,200 ft2 distributed within twelve[unreadable] laboratories and seven offices located centrally in the campus at UTMB. The current scientific staff includes 17 (6[unreadable] Ph.D., 3 M.S., and 6 B.S.) individuals. Specifically, the Proteomics Core will provide established methodologies[unreadable] for sample pre-separation fractionation, 2-dimensional gel electrophoresis (2DE), differential protein staining, gel[unreadable] imaging and analysis, peptide labeling with stable isotopes (iTRAQ) and quantitative mass spectrometry, protein[unreadable] identification by peptide mass fingerprinting via matrix assisted laser desorption ionization time of flight mass[unreadable] spectrometry (MALDI TOF MS), liquid chromatography tandem mass spectrometry (LC/MS/MS), and Luminex[unreadable] multiplex assays for determination of cytokine expression patterns. The specific objectives of the Proteomics Core[unreadable] are: 1. Provide the infrastructure necessary for consistent sample pre-separation fractionation (Project 1); 2.[unreadable] Perform differential protein expression analysis by 2DE and identification of proteins from vascular smooth muscle[unreadable] cells and aortic explant cultures (Projects 1 and 3); 3. Perform differential iTRAQ labeling, LC/MS/MS protein[unreadable] identification of complex protein samples (Projects 1 and 3); and 3. Perform immunoassays of human and mouse[unreadable] chemokines/cytokines in aortic explant cultures from human and mouse samples (Projects 3 and 4). To ensure[unreadable] data quality, validation activities using standard proteins and/ or peptides have been established. Validation of[unreadable] data generated by image analysis with Progenesis software is accomplished utilizing appropriate statistical[unreadable] analyses including Student's t-test for hypothesis and significance testing, Multiple Hypothesis testing corrections,[unreadable] Hierarchical Clustering of control vs. treated, and Analysis of Variance (ANOVA) for time course/dosage[unreadable] dependence of expression. For MS, data quality is ensured through the use of appropriate internal and external[unreadable] standards, while MS instruments are routinely calibrated with external standards. For the Bioplex cytokine[unreadable] measurements, recombinant standards are run with each plate and sample-to-sample variation determined.[unreadable]