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

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Synthesis of Peptides for Use as Immunogens,roducts that are specifically directed against sites of interest, for example, unique regions, highly conserved regions, active sites, extracellular or intracellular domains. Moreover, the ready availability of the pure peptide immunogen against which the antibody was raised means that sera can be r

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Screening of Monoclonal Antibodies Using Antigens Labeled with Acetylcholinesterase,umber of wells to be tested (often a few hundred), and to the small quantities of MAbs available (at best, 300 µL at a few µg/mL), it is not easy at this stage to characterize the fine specificity of the antibodies (i.e., recognition of a precise epitope, inhibitory effect on a biological system, pr

Manifest

Purification of Immunoglobulin G (IgG),tant, or the isolation of a particular antigen binding fraction from such fluids. The former can be achieved by biochemical procedures, whereas the latter usually requires some form of affinity purification.

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Immunohistochemical detection of BromodeoxyurIDine-Labeled Nuclei for In Vivo Cell Kinetic Studies,f tissue by light microscopy. It has also proved extremely valuable for studies in conjunction with flow cytometry and even, for in vivo studies of human tumor cell kinetics (. this vol., Chapter .). We describe here a method to detect dNA synthesis by in vivo labeling of nuclei with BrdU, followed

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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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Electron Microscopic Silver Enhancement for Double Labeling with Antibodies Raised in the Same Specodies (Ab) to be used are raised in the same species, as is usually the case with monoclonal antibodies (MAb), the difficulty arises that the labeled secondary, antispecies Ab used in the first labeling step traps the primary Ab directed against the second antigen, thus leading to a nonspecific signal for the second antigen.

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Epitope Mapping,Monoclonal antibodies (MAbs) are specific immunological tools because they bind to a precise determinant (the epitope) on the surface of a protein. The procedure of identifying the binding site of a MAb is often termed “epitope mapping.”

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