Affinity labeling of antibodies

Antibodies Affinity labeling of antibody combining sites as illustrated by anti-dinitrophenyl antibodies, 46, 479 p-azoben-zenearsonate antibody, 46, 492 affinity cross-linking of heavy and light chains, 46, 501 bivalent affinity labeling haptens in the formation of model immune complexes, 46, 505 DNP-based diazoketones and azides, 46, 508 labeling of antilactose antibody, 46, 516. 

Vol. XLVI [S3]. Affinity Labeling of Antibody Combining Sites as Illustrated by Anti-Dini-trophenyl Antibodies. D. Givol and M. Wilchek. 

Used for affinity labelling of antibodies. Orange or straw needles (pet. ether). Mp 53-55° (52°). 

Affinity Labeling of Antibody Combining Sites as Illustrated by Anti-Dinitrophenyl Antibodies... 

Vn. AFFINITY LABELING OF ANTIBODY MOLECULES OR MYELOMA PROTEINS WITH ANTIBODY ACTIVITY... 

Affinity labeling of immunoglobulins is carried out in a reaction mixture composed of antibody (1 nM), the C aflSnity-labeling reagent (4 fiM), and 0.1 M NaHCOa (pH 9.0). This represents a ratio of labeling rea-... 

Affinity Labeling of Anti-Dnp Antibodies from Various Species by MNBDF - ... 

An important sidelight was the finding that within an individual antibody molecule both sites were either affinity labeled or both were unlabeled (126). Those molecules capable of being bound to a hapten-specific immunoadsorbent after the labeling procedure were essentially free of the label, indicating that few if any molecules were labeled only in one site. Furthermore, antibody molecules absorbed to and eluted from the immunoadsorbent after the initial treatment were resistant to affinity labeling. Thus, antibody molecules appear symmetrical with respect to the capacity of their two active sites to be affinity labeled. 

D. Affinity Labeling of Myeloma Proteins with Antibody Activity... 

Potentially, there are many significant advantages associated with affinity labeling of myeloma proteins, as compared to conventional antibodies. (The myeloma proteins must, of course, have antihapten activity.) The homogeneity of the protein permits detailed sequence analysis and thus a precise localization of the affinity label. In addition, one might expect that in theory all of the sites of the myeloma protein population could be labeled. In the case of conventional antibodies this has generally not been possible because of heterogeneity of active sites evidently not all sites contain an appropriate side chain, or some side chains subject to attack by the affinity label are not sterically available. 

Immunoassays are based on the ability of the immune system to produce a virtually unlimited variety of antibodies each with a high affinity for foreign compounds (immunogens like viruses, bacteria, proteins, and haptens). Analytically, this phenomenon can be exploited by detection of this immunoreaction using labeled antibodies or antigens (i.e., compounds that can be bound by antibodies). The equilibrium between antibody (Ab), antigen (Ag), and immune complex (Ab-Ag) may be expressed as ... 

Pitt JC, Lindemeier J, Habbes HW, Veh RW (1998) Haptenylation of antibodies during affinity purification a novel and convenient procedure to obtain labeled antibodies for quantification and double labeling. Histochem Cell Biol 110 311 322 Polak JM, Van Noorden S (1997) Introduction to immunocytochemistry. BIOS Scientific... 

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