Using the two-dimensional gel electrophoresis and two systems for computer analysis of the resulting maps, we obtained a good quantitative reproducibility of individual spots of silver-stained and Coomassie Blue-stained gels from mouse fetus liver samples. The GelScan-XL system was established for the evaluation of silver-stained gels. The Gel-Image software has shown to be capable of the analy...

In the present study, we compared six different solubilization buffers and optimized two-dimensional electrophoresis (2-DE) conditions for human lymph node proteins. In addition, we developed a simple protocol for 2-D gel storage. Efficient solubilization was obtained with lysis buffers containing (a) 8 M urea, 4% CHAPS (3-[(3-cholamidopropyl) dimethylammonio]-1-propanesulfonate), 40 mM Tris ba...

Serological proteome analysis (SERPA) combines classical proteomic technology with effective separation of cellular protein extracts on two-dimensional gel electrophoresis, western blotting, and identification of the antigenic spot of interest by mass spectrometry. A critical point is related to the antigenic target characterization by mass spectrometry, which depends on the accuracy of the mat...

One third of all genes of various organisms encode membrane proteins, emphasizing their crucial cellular role. However, due to their high hydrophobicity, membrane proteins demonstrate low solubility and a high tendency for aggregation. Indeed, conventional two-dimensional gel electrophoresis (2-DE), a powerful electrophoretic method for the separation of complex protein samples that applies iso...

Background: The numerous methods and techniques employed in proteomics investigations generate vast quantities of heterogeneous data, both results and provenance. A standard representation of proteomics data and techniques is urgently required in order to document a proteomics investigation. A standard representation will facilitate data exchange, verification and further analysis, and pr...

In continuous lines of cultured Drosophila cells two forms of cytoplasmic actin, designated II and III, are detected after a 25- to 120-min pulse label with [35S]methionine. However, only one of these, actin II, accumulates in the cell. With the use of a pulse-chase protocol and two-dimensional gel electrophoresis, it has been found that actin III is synthesized as a precursor of the more stabl...

We have used two-dimensional gel electrophoresis to study the structural transition to the triplex H form of sequences 5'-AAGGGAGAAXGGGGTATAGGGGYAAGAGGGAA-3' where X and Y are any DNA bases. The transition was observed at acid pH under superhelical stress. For X = Y = A or X = Y = G the sequences corresponded to homopurine-homopyrimidine mirror repeats (H-palindrome) which are known to adopt th...

Core histones have been isolated from Schizosaccharomyces pombe and compared electrophoretically to core histones from Saccharomyces cerevisiae and rat liver. The molecular masses of all cognate histones examined were found to be very similar as determined by SDS gel electrophoresis. Histones H3, H2A and H2B from Sch. pombe migrated almost identically to their respective counterparts from S. ce...