Two-dimensional electrophoresis (2-DE) is a powerful and widely used method for the analysis of complex protein mixtures extracted from cells, tissues, or other biological samples. This technique sort’s protein according to two independent properties in two discrete steps: the first-dimension step, isoelectric focusing (IEF), separates proteins according to their isoelectric points (pI); the se...

We utilized two-dimensional gel electrophoresis and immunoblotting (2D-immunoblotting) with anti-Sporothrix schenckii antibodies to identify antigenic proteins in cell wall preparations obtained from the mycelial and yeast-like morphologies of the fungus. Results showed that a 70-kDa glycoprotein (Gp70) was the major antigen detected in the cell wall of both morphologies and that a 60-kDa glyco...

Effective proteome analyses are based on interplay between resolution and detection. It had been claimed that resolution was the main factor limiting the use of two-dimensional gel electrophoresis. Improved protein detection now indicates that this is unlikely to be the case. Using a highly refined protocol, the rat brain proteome was extracted, resolved, and detected. In order to overcome the ...

The synthesis and assembly of vimentin was studied in erythroid cells from 10-d-old chicken embryos. After various periods of [35S]methionine incorporation, cells were lysed in a Triton X-100-containing buffer and separated into a soluble and an insoluble (cytoskeletal) fraction. Analysis of these two fractions by two-dimensional gel electrophoresis shows that vimentin is almost exclusively pre...

The salivary film, denoted the pellicle, formed on oral surfaces is of great importance for oral health and comfort. The present study describes mechanically-assisted sodium dodecyl sulphate (SDS) elution of the in vivo pellicle formed on human enamel and visualisation of the desorbed pellicle proteins using two-dimensional gel electrophoresis (2-DE). To verify this removal of the pellicle, a c...

A method is described which allows to reveal simultaneously the proteolytic patterns of numerous polypeptides separated by two-dimensional electrophoresis. After two-dimensional electrophoresis, the gels were dipped successively in buffers for preequilibration, protease digestion, and reequilibration. They were then returned to the electrophoresis tank, and electrophoresis was continued for a s...

Background: The aim of this study was to compare the sperm protein profile between oligo- and normozoospermic patients with varicocele. Materials and Methods: Sperm samples of 20 normozoospermic and 20 oligozoospermic patients with varicocele were characterized using two dimensional electrophoresis (group 1 and group 2, respectively). Differences in protein expression were established using gel...

The major proteins in isolated synaptic junctions (SJs) and postsynaptic densities (PSDs) have been compared to actin, tubulin, and the major neurofilament (NF) protein by two-dimensional gel electrophoresis and tryptic peptide map analysis. These studies show: (a) tubulin is present in SJ and PSD fractions and is identical to cytoplasmic tubulin, (b) actin in these fractions is very similar to...

Pseudomonas fragi has the ability to grow between 0 and 35 degrees C and grows optimally at 30 degrees C. Cellular proteins from mid-log-phase cells growing from 4 to 34 degrees C were labeled with L-[35S]methionine during 1 generation time and analyzed by two-dimensional gel electrophoresis. The electrophoretic patterns revealed differences in the patterns of protein synthesis over this temper...