Akademik Veri Yönetim Sistemi
Turkish Journal of Analytical Chemistry (Online), cilt.7, sa.3, ss.377-384, 2025 (Hakemli Dergi)
Abstract
Aim: There are various forms of isoelectric focusing. But involving a single glass column and stabilized by the presence of a sucrose gradient, they are preferably used and are the most common pre-application step in 2D electrophoresis. The cylindrical disc-PAGE tube system is possibly not evaluated for isoelectric purposes. The goal here was to try a modified application for the separation of isoform proteins via the disc gel system. And to establish isoform separation and evaluate the implications of this approach for future use.
Methods: Essentially, basic features of disc and isoelectric focusing were integrated. Although conventional disc electrophoresis requires three-layers of gel, but here only one form of gel layer was configured. As a second attempt, the homogenate and ampholite, were deliberately included in the unpolymerised gel medium as if they were gel preparation factors. Then, the separation of the isoform proteins, which were thought to be homogenously distributed in the presence of ampholite, was achieved.
Results: The observation of isoforms of proteins revealed the presence of a distinct pattern of protein bands in their respective isoionic areas. From an analytical perspective, this application exhibited very clear band patterns. The results obtained regarding the isoforms of hemoglobin proteins were looking as informative and descriptive.
Conclusions: The band pattern observed in the present approach clearly showed multiple isoform characters and simply demonstrated the differences of this novel application. It can be used to analysis and separate multiple of proteins in the first dimension of conventional two-dimensional electrophoresis.
Keywords:
IEF, IPG-Dalt, ampholite, isoform proteins, hemoglobins multiplicity, disc-PAGE System