免疫学技术

Cell kinetics is defined as the measurement of time parameters in biological systems. Traditionally, this has involved the use of radioactive precursors of DNA, such as tritiated thymidine (3HTdR), and autoradiography to detect their incorporation into DNA. This technique has provid ...

The detection and quantification of cells undergoing proliferation have centered on methods that identify specific stages of the cell cycle. Thus, the replication of DNA or S-phase (the labeling index) is detected by the use of -thymidine and autoradiography, or by bromodeoxyuridine and ...

One of the most common methods in immunohistochemistry involves the use of an antibody to the antigen of interest detected indirectly with an enzymelabeled antispecies secondary antibody. The enzyme catalyzes the formation of a colored insoluble reaction product at the antigen site. It ...

As long ago as 1857 (1), Michael Faraday in an address to the Royal Society recognized the reactive properties of colloidal metal particles (in particular gold) with light. Since 1933 (2), a range of methods have been devised for producing gold sols. It was not until 1971, however, that Faulk and Taylor (3) pu ...

Highly specific antibodies directed against minor proteins, present in small amounts in biological fluids, or against nonsoluble cytoplasmic or membranous proteins, are often difficult to obtain. The main reasons for this are the small amounts of protein available after the various c ...

All immunocytochemical techniques are based on the same principle of incubating the target antigen with an appropriate antibody solution, but they may incorporate one from a range of different microscopically dense markers for visualizing, the sites of binding. Light microscope im ...

Electron microscopy permits the detailed study of cell relationships within tissues and organelles within cells. Two ultrastructural techniques in which gold probes have proven invaluable are immunocytochemistry and in situ hybridization, and these will be described here. Un ...

In immunoelectron microscopy (IEM), simultaneous labeling of two or more antigens on the same section is desirable for many applications. If the antibodies (Ab) to be used are raised in the same species, as is usually the case with monoclonal antibodies (MAb), the difficulty arises that the label ...

Analysis of cell surface antigen density and, thus, the distribution of cell surface molecules among individual members of complex cell populations can be achieved rapidly and accurately by flow cytometry. This approach has been revolutionized over the last 20 yr by the codevelopment of m ...

As indicated in Chapter 33, flow cytometry has developed rapidly to provide a powerful means of characterizing complex cell populations, both in terms of quantitative analysis of functional cell-associated molecules, and, as is considered here in more detail, the simultaneous analys ...

Apoptosis is a physiological, programmed mode of cell death, which is necessary for tissue modeling and organogenesis in embryonic development and in the control of homeostasis in a diversity of tissue types (1). The distinct morphological features of apoptosis clearly distinguish it f ...

Immunomagnetic beads are uniform, polymer particles coated with a polystyrene shell that provides both a smooth, hydrophobic surface to facilitate absorption of molecules, such as antibodies, and surface hydroxyl groups that allow covalent chemical binding of other bioreactive ...

The rapid evolution of monoclonal antibody (MAb) technology has resulted in an ever-growing repertoire of proteins becoming accessible for study using immunohistochemical techniques. Nuclear antigens represent a class of proteins of increasing significance and diversity; ...

Molecular hybridization is a useful technique for identifying specific target sequences even when they are present as a single copy in a complex population of highly heterogeneous gene sequences. It can be performed either on a solid matrix on which pure DNA (or RNA) is bound (blot hybridizatio ...

In situ hybridization may be defined as the detection of nucleic acids in situ in cells, tissues, chromosomes, and isolated cell organelles. The technique was described in 1969 by two separate groups who demonstrated repetitive ribosomal sequences in nuclei of Xenopus oocytes using radio ...

Immunization protocols vary greatly among laboratories. In general, there are no hard and fast rules, and most protocols give satisfactory results. The methods described below are designed to give optimal results with minimal injury to the test animal, and we have used them extensively and s ...

Conjugates of antibodies or antibody fragments with foreign proteins have attracted interest because of their therapeutic potential as antitumor agents in vivo. Localization of the complex to the target site is dependent on the specificity of the antibody moiety, and local cytotoxic a ...

With the advent of nonradioactive probes during the past decade, in situ hybridization has become an immensely valuable tool in the hands of: 1. Developmental biologists and cell biologists for the detection of mRNA in cells and tissue sections (1

Much of our current understanding of the molecular details of the activity and interactions of proteins has stemmed from the ability to isolate cDNA encoding these proteins. Computer-based analysis of deduced amino acid sequence allows predictions to be made about their structure and fu ...

Recombinant DNA technology provides a means to mimic nature and build multidomain proteins that display several functions. This ability is being applied by the pharmaceutical industry to develop novel products. Fusion proteins of antibodies and enzymes formed one of the earliest exa ...