Titlebook: Immunochemical Protocols; Margaret M. Manson Book 19921st edition Humana Press 1992

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Raising Polyclonal Antibodies Using Nitrocellulose-Bound Antigen,neous proteins, are often difficult to obtain. The main reasons for this are the small amounts of protein available after the various classical purification processes and the low purity of the proteins.

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Production and Characterization of Antibodies Against Synthetic Peptides,he methods described below are designed to give optimal results with minimal injury to the test animal, and we have used them extensively and successfully for several years (.–.). Peptide immunizations differ from those in which the immunogen is a larger macromolecule in that maximal antipeptide tit

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Screening of Monoclonal Antibodies Using Antigens Labeled with Acetylcholinesterase,öhler and Milstein (.). These workers have shown that lymphocytes can be immortalized and subsequently cultured after somatic fusion with genetically selected myeloma cells. Usually, once fusion between spleen cells and myeloma cells has been performed, cells are suspended in a large volume of selec

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Purification of Immunoglobulin G (IgG),re supernatant containing monoclonal antibodies (MAbs). However, a much “cleaner” result can often be obtained if some form of enrichment or isolation of immunoglobulin is employed. Some procedures, such as conjugation with isotopes, fluorochromes, or enzymes, and preparation of immunoaffinity colum

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Enzyme-Antienzyme Method for Immunohistochemistry,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. The advantage of an enzyme label is that the nuclei can be counterstained, thereby revealing the tissue architecture, and that the stain fades slowly, if

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,Avidin–Biotin Technology, application (for general reviews, . refs. 1 and 2). For example, a fluorescent form of avidin can be used for fluorescence microscopy, fluorescence-activated cell sorting, and in some cases, for immunoassay. Like-wise, an avidin-enzyme conjugate can be used for immunoblotting, immunoassay, light mi