Comparison of First-dimension Separations for 'MudPIT' Proteomic Studies on Endometrial Tissue

摘要

From previous 2D gel experiments, we knew that endometrial tissue contained a large quantity of plasma proteins. Because these proteins were denatured during the extraction process, they could not be efficiently removed by the antibody-based depletion method. When digestion is performed prior to the first-dimension separation, albumin peptides (for example) are distributed throughout all of the fractions, potentially causing interferences in every LC/MS/MS analysis. The purpose of this study was to determine if separation at the protein level (the geLC method) might "concentrate" these interfering proteins into a few gel slices (i.e., a few LC/MS/MS fractions), allowing the detection of lower-abundance proteins in the remaining fractions.