基因表达差异显示实验

Radiolabeling is a highly sensitive method for protein detection, which is easily performed by the incorporation of radioactive amino acids into proteins. This makes radiolabeling a method of choice for visualizing proteins separated on two-dimensional (2-D) gels. This chapter pre ...

Heparins are negatively charged polydispersed linear polysaccharides which have the ability to bind a wide range of biomolecules including enzymes, serine protease inhibitors, growth factors, extracellular matrix proteins, DNA modification enzymes and hormone receptor ...

It is widely known that the progress of proteomics mostly depends on the development of more advanced and sensitive protein separation technologies. Immobilized metal affinity chromatography (IMAC) is a useful protein fractionation method used to enrich metal associated protei ...

Chromatofocusing has many potential applications in the field of proteomics, such as for the isolation and removal of major sample components to facilitate the analysis of low-abundance components, and for sample prefractionation prior to a subsequent separation using SDS-PAGE, n ...

High-resolution 2-dimensional gel electrophoresis (2DGE) is a key technology in the analysis of cellular proteomes particularly in the field of microbiology. However, the restricted resolution of 2DGE and the limited dynamic range of established staining methods limit its usefu ...

Proteome analysis by two-dimensional gel electrophoresis (2-DGE) faces significant challenges because of the complexity of biological samples. However, the complexity of a protein sample can be reduced prior to 2-DGE by applying protein fractionation. Protein fractionation a ...

Because the dynamic range of most cell or tissue proteomes is enormous, separation of such complex protein samples by two-dimensional electrophoresis (2-DE) on broad pH gradients often results in the visualization of only the most abundantly expressed proteins. It is, therefore, often b ...

Reproducible techniques for the prefractionation of proteins prior to two-dimensional gel electrophoresis (2-DE) are essential for increasing the number of unique proteins that can be identified and assayed following biological experimentation. A simple and robust techniq ...

Quantitative proteomics has become a pivotal tool that has been applied to the investigation of many different biological processes such diverse as the detection of biomarkers in tissue samples, the regulation of cell signaling, and the characterization of protein interactions. St ...

Isotope-coded two-dimensional maps, with either D/D-acrylamide or D/D 2-vinyl pyridine, are described in detail. They have the advantage of running the two samples under investigation within a single slab gel, thus minimizing errors because of spot matching with software packages when s ...

Apoptotic and programmed cell death are characterized by, and indeed were first discovered from observations of, remarkable morphological changes that occur in the nucleus (see 1 for a comprehensive review of apoptosis and programmed cell death). Thus, light and electron microscopy w ...

The study of DNA damage holds a wide interest within both basic and applied fields of research. Elucidating the mechanisms involved in the generation of DNA damage, and the consequences of this damage, will have an enormous impact on multiple fields of scientific research and will ultimately le ...

A number of exogenous and endogenous toxic agents may damage DNA, leading to genomic instability and transcriptional infidelity. Genetic or acquired defects in DNA repair mechanisms also contribute to exacerbate DNA damage. Progressive accumulation of DNA injury may alter the gene ...

Apoptosis is a widespread phenomenon, which plays an important role in many physiological events as well as pathological processes (1). It was originally defined by its unique ultrastructural features, which were detected by electron microscopy (2,3). They are cytoplasmic shrinkage, ...

The initial concept of apoptosis was proposed by Kerr et al. (1972) (1) and was based on characteristic morphological criteria. Transmission electron microscopy (TEM) allows visualization of fine details of cellular morphology and is one of the most reliable methods for detecting of apop ...

The presence of a multitude of DNA strand breaks resulting from fragmentation of nuclear DNA by the caspase-activated DNase is one of the most chaacteristic features of apoptotic cells (1,2). A widely used methodology to detect apoptotic cells, thus, relies on labeling DNA strand breaks in situ, ...

The nick translation procedure has been introduced in 1977 by Righy et al. (1). It is commonly used to label purified DNA in vitro. This technique relies on combined 5′→3′ polymerase and 5′→3′ exonuclease activities of Escherichia coli DNA polymerase I. If a nick or single strand break with a 3′ hydroxyl term ...

The Klenow fragment of DNA polymerase I, first described by Klenow and Henningson in 1970 (1), consisted of a 75 kDa proteolytic fragment of the Kornberg polymerase of Escherichia coli. Klenow is now usually obtained as a recombinant protein expressed from a truncated polA gene (2). To polymerize t ...

Mainly as a result of the booming interest in apoptosis and of the discovery of the occurrence of massive DNA breaks as a major feature of this type of cell death, during the past few years polymerase-based assays for detection of DNA breaks in cellular nuclei have found worldwide application in many fie ...

In 1992 Gavrieli et al. published a seminal article showing that apoptotic cells could be detected by an in situ assay (1). The labeling method depends on the ability of terminal deoxyribonucleotidyl transferase to add nucleotides to breaks in DNA, and has generally been termed TUNEL (terminal d ...