Two-dimensional electrophoresis in basic and clinical research, as exemplified by studies on the exocrine pancreas.
I summarize our experience in the development and use of two-dimensional isoelectric focusing/SDS gel electrophoresis for the study of soluble proteins, in particular those of the exocrine pancreas. The importance of adding protease inhibitors to the first-dimension isoelectric focusing gel to prevent the autoactivation of potential pancreatic proteases and subsequent proteolysis is discussed. Recently developed methods for both the indirect and direct identification of protein spots by biological activity are summarized. I give examples of the importance of this technique in the study of basic mechanisms involved in the synthesis, segregation, and secretion of exportable proteins in the exocrine pancreas, as well as in the search for biochemical markers of pancreatic disease.
D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D007525:Isoelectric Focusing; D007526:Isoelectric Point; D008968:Molecular Conformation; D010179:Pancreas / Q000201:enzymology / Q000378:metabolism*; D010182:Pancreatic Diseases / Q000378:metabolism*; D010189:Pancreatic Juice / Q000032:analysis*; D011480:Protease Inhibitors; D014176:Protein Biosynthesis; D011506:Proteins / Q000032:analysis* / Q000235:genetics
D011480:Protease Inhibitors; D011506:Proteins