ANTIBODY ENZYME CONJUGATES THE PREPARATION OF INTERMOLECULAR CONJUGATES OF HORSERADISH PEROXIDASE AND ANTIBODY AND THEIR USE IN IMMUNOHISTOLOGY OF RENAL CORTEX

The studies reported were done with the objectives of preparing several different peroxidase-labeled antibody conjugates and of testing their usefulness for the detection of soluble and insoluble tissue antigens. Glutaraldehyde, toluene diisocyanate and N,N'-dicyclohexyl carbodiimide were used...

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Bibliographic Details
Published in:The journal of histochemistry and cytochemistry 1973-03, Vol.21 (3), p.233-240
Main Authors: CLYNE, DAVID H, NORRIS, STEPHEN H, MODESTO, ROSARIO R, PESCE, AMADEO J, POLLAK, VICTOR E
Format: Article
Language:English
Subjects:
Animals
Antibodies - analysis
Antigen-Antibody Reactions
Centrifugation, Density Gradient
Chromatography, Gel
Fibrinogen
gamma-Globulins
Goats - immunology
Haplorhini
Histocytochemistry
Humans
Immunoelectrophoresis
Iodine Isotopes
Kidney - anatomy & histology
Kidney - immunology
Kidney Cortex - immunology
Kidney Diseases - immunology
Peroxidases - analysis
Plants - enzymology
Rabbits - immunology
Schiff Bases
Solubility
Staining and Labeling
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Summary:The studies reported were done with the objectives of preparing several different peroxidase-labeled antibody conjugates and of testing their usefulness for the detection of soluble and insoluble tissue antigens. Glutaraldehyde, toluene diisocyanate and N,N'-dicyclohexyl carbodiimide were used to cross-link horseradish peroxidase to goat anti-rabbit immunoglobulin G; the resulting conjugates were characterized by molecular size and enzymatic and immunologic activity. They were then tested for their properties as immunohistologic reagents using 0.5-µ sections of freeze-substituted paraffin-embedded renal cortical tissue. Excellent results were obtained with a highly polymerized conjugate made with glutaraldehyde and with an unpolymerized conjugate made with toluene diisocyanate. With the use of these conjugates tissue localization of both soluble and insoluble antigens was achieved after subsequent fixation of tissue and counterstaining with periodic acid-Schiff-hematoxylin.
ISSN:0022-1554
1551-5044
DOI:10.1177/21.3.233