抗体抗原实验

The application of in situ hybridization and the underlying rationale are described in Chapter 46. In situ hybridization of nucleic acid probes to cellular targets of biological specimens affixed to a slide has benefited greatly from the concepts and strategies so elegantly developed u ...

Immunohistochemical procedures have become an integral part of the clinical laboratory routine, evolving from a research tool to a diagnostic necessity in pathology. As a specifically defined laboratory section, the Immunohistochemistry Laboratory must meet federally man ...

In situ polymerase chain reaction (PCR) is a very powerful tool, which enhances our ability to detect minute quantities of a rare, single copy number, target nucleic acid sequences in freshly frozen or paraffin-embedded intact cells or tissue sections (1–10). In 1986, the introduction of PCR met ...

In 1974, the presence of intracytoplasmic immunoglobulins in plasma cells was demonstrated in routinely processed paraffin tissue (the mainstay of diagnostic pathology) using an immunoperoxidase technique (1). This new capability has introduced the surgical pathologist to a t ...

Several immunological procedures can be successfully carried out using nonpurified antibodies, such as unfractionated antisera or ascitic fluid/culture supernatant containing monoclonal antibodies (MAbs). However, a much “cleaner” result can often be obtained if some form ...

Hybridoma technology has made possible the production of specific homogeneous antibodies (monoclonal antibodies ) with predefined binding characteristics that can be produced in large amounts from immortal cell lines. MAbs can be exquisitely specific, but preparations cont ...

Bispecific antibodies (BsAbs) are hybrid molecules designed to combine two antibodies with different specificities. This dual specificity has led to their application in a variety of diagnostic situations that require the crosslinking of two protein surfaces, for example, antig ...

Cytotoxic antibody—toxin conjugates made using antibodies and ribosome-inactivating proteins (RIPs) are prepared using chemical crosslinking methods (1,2 and Chapter 13). Gel permeation chromatography is used as a first step to purify conjugate molecules from the reaction mi ...

Conjugates of antibodies with plant toxins, such as ncin and abrin, are potent cytotoxic agents that selectively eliminate target cells from mixed cell cultures in vitro, and have great promise as antitumor agents in cancer therapy (1). Ricin and abrin are protein toxins consisting of two diff ...

Chemiluminescence results from reactions with a very high energy yield, which produce a potentially fluorescent product molecule; reaction energy passed to the product may result in an excited state and subsequent production of a single photon of light. The light yield is usually low, but can ...

All immunochemical procedures require a suitable antiserum or monoclonal antibody raised against the antigen of interest. Polyclonal antibodies are raised by injecting an immunogen into an animal and, after an appropriate time, collecting the blood fraction containing the anti ...

The ability to detect specific proteins that have been immobilized on nitrocellulose or polyvinylidene difluoride (PVDF) membranes has become an invaluable technique in all areas of the biological sciences. In the past, visualization of low-abundance proteins required the use of ra ...

Proteins, blotted from polyacrylamide gels onto nitrocellulose sheets (Western blots) can be stained nonspecifically with a variety of dyes, or they can be identified individually by probing with appropriate antibodies. These procedures may be performed on duplicate blots, stai ...

Western blotting (reviewed in refs. 1–3) refers to formation and detection of an antibody—antigen complex between an antibody and a polypeptide that is immobilized on denvatized paper. Most commonly, polypeptides in a complex mixture are separated by electrophoresis through polyac ...

Immunohistochemical stains use antibodies to identify specific constituents in tissue sections. In order to detect the site of reaction, the antibody is labeled with an enzyme that can be reacted with a suitable substrate to give a colored product. The alternative is to use a fluorescent label. ...

This chapter extends the use of the technique described in Chapter 21 to two-dimensional (2D) protein gels, as well as containing some alternatives and modifications to the method.

The classical technique for identifying cells engaged in DNA synthesis is by their uptake of -thymidine, detected using autoradiography. However, this method can be inconvenient, as specialized darkroom and radioisotope facilities are required, with the potential health hazard ...

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 ...