细胞技术

This chapter covers the breakdown of the process of angiogenesis into simple assays to measure discrete endothelial cell functions. The techniques described are suitable for studying stimulators or inhibitors of angiogenesis and determining which aspect of the process is modula ...

Tissue blood flow rate (F) is a critical parameter for assessing functional efficiency of a blood vessel network following angiogenesis. This chapter aims to provide the principles behind estimation of F and a practical approach to its determination in laboratory animals using small, re ...

The host response observed after the application of an appropriate stimulus, such as mechanical injury or injection of neoplastic or normal tissue implants, has allowed the cataloguing of a number of molecules and cells involved in the vascularization of normal repair or neoplastic tiss ...

The search for rapid and reproducible in vivo angiogenesis and antiangiogenesis assays is an area of intense interest. These types of assays are extremely useful in testing putative drugs and biological agents and for the comparison and enhancement of in vitro tests. The Matrigel plug assay is ...

Intravital microscopy represents an internationally accepted and sophisticated experimental method to study angiogenesis, microcirculation, and many other parameters in a wide variety of neoplastic and nonneoplastic tissues. Since 1924, when the first transparent cham ...

Continuous monitoring of neovascular growth in vivo is required for the development and evaluation of drugs acting as suppressors or stimulators of angiogenesis. The cornea assay consists of the placement of an angiogenesis stimulus (tumor tissue, cell suspension, growth factor) in ...

In vivo preclinical assays are required to screen potential agents that target the tumor vasculature. Here, a hollow fibre-based assay for the quantification of neovasculature in the presence or absence of an agent that potentially targets tumor neovasculature is described. The neova ...

Reactive oxygen species (ROS) are constantly produced by skeletal muscle and this production is increased during contractile activity. Understanding the role that ROS play in skeletal muscle requires an understanding of the species of ROS produced, the subcellular site of producti ...

Matrix metalloproteinases (MMPs) are a family of zinc-dependent proteinases associated with extracellular matrix degradation, cellular migration, tissue remodeling, and angiogenesis. The activity of MMPs is regulated by the tissue inhibitors of metalloproteinases (TI ...

Cholesterol-rich microdomains present on the plasma membrane appear to play an important role in spatio-temporal regulation of cell signaling and cell adhesion processes. Compositional heterogeneity of these microdomains and their coalescence during cell–cell interact ...

Dye affinity chromatography is a purification technique offering unique selectivities and high purification factors. Dye ligands may act as substrate analogs, offering affinity interactions with their corresponding enzymes. This chapter describes a dye ligand chromatog ...

Displacement chromatography has several advantages over the nonlinear elution technique, as well as the linear elution mode, such as the recovery of purified components at high concentrations, less tailing during elution, high throughput and high resolution. Displacer affinity ...

Immobilized Metal ion affinity chromatography (IMAC) is a ubiquitous technique in Modern recombinant production and purification. The wide range of expression vectors for the production of histidine-tagged recombinant proteins as well as the variety of stationary supports for ...

The HQ (H = histidine, Q = glutamine) tag is a small fusion tag that can be isolated using immobilized metal affinity columns. HQ-tagged proteins can be expressed and purified from bacterial cells under native and denaturing conditions, mammalian cells, insect cells, wheat germ and rabbit retic ...

Maltose-binding protein (MBP) is a carrier protein for high level recombinant protein and peptide production from either the cytoplasm or periplasm of Escherichia coli. The affinity matrix for purifying MBP-passenger proteins utilizes amylose covalently attached to magnetic b ...

Where an affinity tag has served its purpose, it may become desirable to remove it from the protein of interest. This chapter describes the removal of such fusion partners from the intended protein product by cleavage with site-specific endoproteases. Methods to achieve proteolytic cleav ...

HaloTag™ is a protein fusion tag which was genetically engineered to covalently bind a series of specific synthetic ligands. All ligands carry two groups, the reactive group and the functional/reporter group. The reactive group, the choloroalkane, is the same in all the ligands and is involved ...

The use of affinity tags and especially histidine tags (His-tags) has become widespread in molecular biology for the efficient purification of recombinant proteins. In some cases, the presence of the affinity tag in the recombinant protein is unwanted or may represent a disadvantage for the ...

Synthetic affinity ligands can circumvent the drawbacks of natural immunoglobulin (Ig)-binding proteins by imparting resistance to chemical and biochemical degradation and to in situ sterilization, as well as ease and low cost of production. Protein L (PpL), isolated from Peptostr ...

Large repertoires of peptides displayed on bacteriophage have been extensively used to select for ligand-binding molecules. This is a relatively straightforward process involving several cycles of selection against target molecules, and the resulting ligands can be tailored ...