The oxidation and reduction of cysteine residues is emerging as an important post-translational control of protein function. We describe a method for fluorescent labelling of either reduced or oxidized thiols in combination with two-dimensional sodium dodecyl sulphate polyac ...
Oxidative stress has been shown to be one of the mechanisms involved in a number of diseases, including neurodegenerative disorders, ischemia, cancer, etc. Oxidative stress occurs mainly due to an imbalance between oxidant and antioxidant systems. Oxidants can damage virtually all bio ...
Silver staining detects proteins after electrophoretic separation on polyacrylamide gels. Its main positive features are its excellent sensitivity (in the low nanogram range) and the use of very simple and cheap equipment and chemicals. The sequential phases of silver staining are p ...
Our two-dimensional gel electrophoresis (2DE) protocol has been continuously improved in our laboratory since its inception 30 years ago. An updated version is presented here. This protocol is a result of our experience in proteome analysis of tissue extracts, cultured cells (mammali ...
Two-dimensional gel electrophoresis (2DE) separates proteins by molecular charge and molecular size. Proteins are first solubilised in a denaturing buffer containing a neutral chaotrope, a zwitterionic or neutral detergent, and a reducing agent. First-dimension isoelectr ...
A state of oxidative stress (OS) can occur when there is an imbalance between the rate of reactive oxygen species (ROS) production and their detoxification. Under OS conditions sulphur-containing residues are particularly susceptible to oxidation, and this can result in transient for ...
Proteomics often involves systematic analyses of proteomes that are constantly changing in response to changes in the environment of the cell, tissue, or organism being analyzed. Due to limitations of all current protein profiling methods, powerful, reliable proteome prefractio ...
Proteomic profiling of subcellular compartments has many advantages over traditional proteomic approaches using whole cell lysates as it allows for detailed proteome analysis of a specific organelle and corresponding functional characteristics. The microsome is a critic ...
Two-dimensional electrophoresis (2DE) is one of the most effective methods for the reliable separation of proteins in a single gel. In our proteome analyses of human chromosomes, we used two types of 2DE: two-dimensional isoelectric focusing SDS–polyacrylamide gel electrophoresis ...
The importance of the mitochondrion in maintaining normal cellular physiology has long been appreciated. Recently there has been an upsurge in mitochondrial research due to increased recognition that a number of diseases are caused by defective functioning of this key intracellul ...
About 30 years ago two-dimensional gel electrophoresis (2DE) was developed independently by Klose and O’Farrell representing the combination of two orthogonal separation techniques. In the first dimension the proteins are separated by isoelectric focusing (IEF) according to t ...
Most proteomic analyses require prefractionation and protein purification strategies to achieve maximal proteome coverage, especially in plants in which cells often have a few highly abundant proteins and substances like polyphenols or secondary metabolites that can have si ...
Application of the two-dimensional gel electrophoresis (2DE) protocols which were developed for samples of mammalian origin gives unsatisfactory results when used in samples from marine molluscs. This chapter describes a detailed protocol of 2DE that can be applied to these organi ...
Our protein extraction protocol for two-dimensional gel electrophoresis (2DE) was updated to meet current needs in the field of proteomics. This protocol summarizes our experience using this method since its introduction over 30 years ago. We provide a total as well as fractionated extra ...
Proteomic studies of the free-living nematode Caenorhabditis elegans have recently received great attention because this animal model is a useful platform for the in vivo study of various biological problems relevant to human disease. In general, proteomic analysis is carried out in o ...
Protein expression profiling is defined in general as identifying the proteins expressed in a particular tissue, under a specified set of conditions and at a particular time, usually compared to expression in reference samples. This information is useful in drug discovery and diagnosis ...
Protein solubilization for two-dimensional electrophoresis (2DE) has to break molecular interactions to separate the biological contents of the material of interest into isolated and intact polypeptides. This must be carried out in conditions compatible with the first dimens ...
The introduction of two-dimensional fluorescent difference gel electrophoresis has enabled the extensive screening of differential protein expression levels with higher confidence and greater sensitivity than using the classical two-dimensional electrophoresis ...
The overall study of post-translational modifications (PTMs) of proteins is gaining strong interest. Beside phosphorylation and glycosylation, truncations of the nascent polypeptide chain at the N- or C-terminus are by far the most common types of PTMs. Nevertheless, little attenti ...
Cell kinetics is a science that has as its objective an understanding of how cells mature with time and how they respond to outside influences (e.g., exposure to radiation and drugs). Emphasis is placed on the rates at which these events occur. Consider, for example, the application of cell cycle kineti ...
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