Tim Courtney, Chemical Engineering
"Evaluation of Protein Quantitation Assays for Use with Two Dimensional Gel
Faculty Mentor: Dr. Mark R. Marten
Expected Graduation Date: Spring 2009
Two Dimensional Gel Electrophoresis (2DE) is used to separate complex mixtures of proteins for further analysis via mass spectrometry. Before a protein mixture can be separated using 2DE, it is critical to know the total protein concentration; using too much or too little protein will not yield useful information. Protein quantitation assays allow us to determine the concentration of a protein sample; however, the detergents and reducing agents necessary for 2DE interfere with common assays. A variety of specialized protein assay kits (e.g., 2D-Quant, RC-DC, and EZQ) have been developed to overcome this problem, but proteomics results from the Marten lab have shown that results depend on which assay is used, and there is no available literature comparing the efficacy of these assays. My goal is to analyze results from a variety of 2DE compatible assays and recommend the best assay in terms of accuracy, reproducibility, and ease of use. Utilizing the best assay available will benefit proteomics research in the Marten lab and in the entire proteomics community.