2D-PAGE based poroteomics and protein analysis – unravelling biological networks – detailing individual protein function

抄録

Whatever the wealth of information buried in the genome of organisms is converted to in order to create functional protein networks - the perfectly organized interaction of biologic macromolecules is of stunning complexity. Several lines of development of protein analytical technology have evolved as todays tools towards dissecting functional interactions of proteins. Though mass spectrometry, liquid chromatography, protein chips and other techniques have gained widespread acceptance as powerful sources of information, 2DE-PAGE, the electrophoretic separation of complex protein mixtures in two different dimensions, retains its extraordinary position. The unequaled power of separation and the unique feature of simultaneous characterization of thousands of discrete protein entities underline the importance of this method. We have been specializing in the 2DE-PAGE variant acc. to Klose and Kobalz, called NEPHGE (non-equilibrium pH-gradient electrophoresis) for more than a decade. Ever since we work to improve and optimize this method. We have developed proprietary formulations of gel and sample preparation solutions as well as introducing newly designed equipment. With our current state of the art technology we prepare up to 40 by 30 cm giant gels with more than 10,000 resolved protein spots. In combination with NEPHGEs intrinsic advantages of high sample load capacity, the superior ability to resolve basic proteins and versatility in view of subsequent analytical and micro-preparative options, this platform is ideal for many of todays proteomics objectives. Embedded in a wide range of assistant protein analytical techniques, like adjunct electrophoretic methods, MALDI-MS as well as LC ESI-MS/MS characterization of proteins, automated Edman micro sequencing and reference peptide synthesis, we are well equipped to describe complicated biological situations and to answer protein analytical questions.