Differential In-Gel Electrophoresis in a High-Throughput Environment
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Proteomics deals with the large-scale analysis of proteins expressed by the genome of an organism. In general, the aim is to search for qualitative and quantitative changes in protein expression that occur as a function of development, disease, environmental insults, or treatment. Because of the complexity of the proteome of most organisms, the complete and rapid mapping of protein expression changes and characterization of posttranslational modifications is challenging and will require an analytical effort that exceeds the technology and throughput that exist in most standard biochemistry laboratories (1 While novel automated methods for rapid separation and analysis of proteins are being evaluated in many laboratories, it is widely accepted that twodimensional gel electrophoresis (2-DE) is still the benchmark for large-scale separation of complex protein mixtures. The availability of standard reagents and equipment, and development of many modifications of the 2-DE process, have led to remarkable improvements in reproducibility (2,3). However, it is still difficult to fully duplicate the pattern of protein expression using conventional 2-DE methods (4,5), particularly when conducting multiple comparative and quantitative proteomics experiments in a high-throughput research environment where reproducibility and speed are critically important parameters.
KeywordsSodium Dodecyl Sulfate Protein Spot Equilibration Buffer Picking List Spot Picker
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