Localization of ?-d-glucosyl and ?-d-mannosyl groups of mucosubstances with concanavalin A and horseradish peroxidase

1975 ◽  
Vol 44 (1) ◽  
pp. 39-45 ◽  
Author(s):  
J. A. Kiernan
Keyword(s):  
Horseradish Peroxidase ◽  
Concanavalin A

Concanavalin A-horseradish peroxidase bridge staining of alpha-1 glycoproteins separated by isoelectric focusing on polyacrylamide gel.

1976 ◽  
Vol 24 (8) ◽  
pp. 908-914 ◽  
Author(s):  
R C Allen ◽  
S S Spicer ◽  
D Zehr
Keyword(s):  
Horseradish Peroxidase ◽  
Isoelectric Focusing ◽  
Concanavalin A ◽  
Periodic Acid ◽  
Polyacrylamide Gel ◽  
New Method ◽  
Polyacrylamide Gels ◽  
Periodic Acid Schiff ◽  
Coomassie Blue

The Coomassie Blue protein stain and the periodic acid-Schiff stain for glycoproteins are compared to a new method of staining glycoproteins resolved electrophoretically. The method utilizes a Concanavalin A-horseradish peroxidase sequence to visualize selectively glycoproteins with terminal or internal mannose or terminal N-acetylglucosamine. The method applied to characterization of M and Z allele products of alpha-l-antitrypsins separated by isoelectric focusing of polyacrylamide gels slabs have revealed differences in carbohydrate content of various components that were previously undetected.

A Mediator-free Horseradish Peroxidase Biosensor Based on Concanavalin A

2006 ◽  
Vol 34 (3) ◽  
pp. 399-403 ◽  
Author(s):  
Mengmeng Guo ◽  
Yunhui Yang ◽  
Zhijie Wang ◽  
Guoli Shen ◽  
Ruqin Yu
Keyword(s):  
Horseradish Peroxidase ◽  
Concanavalin A

scholarly journals Histochemical differentiation of complex carbohydrates with variants of the concanavalin A-horseradish peroxidase method.

1978 ◽  
Vol 26 (4) ◽  
pp. 233-250 ◽  
Author(s):  
T Katsuyama ◽  
S S Spicer
Keyword(s):  
Horseradish Peroxidase ◽  
Concanavalin A ◽  
Periodate Oxidation ◽  
Liver Glycogen ◽  
Class Ii ◽  
Class Iii ◽  
Horseradish Peroxidase Method ◽  
Hrp Method ◽  
Complex Carbohydrates ◽  
Mucous Neck Cells

Various treatments carried out prior to the concanavalin A-horseradish perioxidase (HRP) method have been found to affect the staining and have permitted differentiation of three main classes of complex carbohydrates in the rat alimentary tract. Class I mucosubstances lose and class II and III paradoxically gain concanavalin A-horseradish peroxidase reactivity after periodate oxidation. Class II mucosubstances lose whereas class III retain or increase their reactivity with a reduction step interposed between oxidation and concanavalin A-horseradish peroxidase staining. Mucous neck cells, pyloric glands, Brunner's glands and mast cells exhibit strong class III staining, whereas other sites such as intestinal goblet and salivary gland acini differ widely in their type of staining. Liver glycogen stains like mucosubstances in an unstable subgroup of class III. The paradoxical increase in concanavalin A binding during oxidation correlates with the appearance of Schiff reactivity implicating oxidation of vicinal hydroxyls as the basis for the effect. The periodate-induced staining is therefore, thought to result from an oxidative disruption of linkages between vicinal hydroxyls of neighboring sugars and hydroxyls of mannose required for concanavalin A binding. Staining with the described concanavalin A-horseradish peroxidase variants appears to afford information concerning cytochemical distribution of mannose-rich glycoproteins as well as differences among these substances in the relation of mannose to neighboring sugars.

scholarly journals Adsorption of horseradish peroxidase, ovomucoid and anti-immunoglobulin to colloidal gold for the indirect detection of concanavalin A, wheat germ agglutinin and goat anti-human immunoglobulin G on cell surfaces at the electron microscopic level: a new method, theory and application.

1977 ◽  
Vol 25 (11) ◽  
pp. 1187-1200 ◽  
Author(s):  
W D Geoghegan ◽  
G A Ackerman
Keyword(s):  
Horseradish Peroxidase ◽  
Concanavalin A ◽  
Colloidal Gold ◽  
Wheat Germ Agglutinin ◽  
Wheat Germ ◽  
Indirect Detection ◽  
Electron Microscopic Level ◽  
Specific Cell ◽  
Major Protein ◽  
Electron Microscopic

A method is described for the adsorption of selected macromolecules to colloidal gold which is then used as an electron dense marker for the indirect detection of specific cell surface molecules. Membrane bound concanavalin A, which binds specific sugars on horseradish peroxidase, and wheat germ agglutinin, which binds specific sugars on ovomucoid are detected indirectly with gold labeled horseradish peroxidase and ovomucoid, respectively. Goat anti-human IgM on blood lymphocytes is detected with gold labeled rabbit anti-goat IgG. In the preparation of colloidal gold labeled proteins, the problems of flocculation of colloidal gold by proteins and nonadsorption of proteins to colloidal gold, are solved through a combination of concentration of protein and pH variable adsorption isotherms, which allows one to determine the conditions for adsorption of proteins to colloidal gold. Adsorption is pH dependent, the pH conditions correlating with the isoelectric point(s) of the major protein fraction(s); adsorption is influenced by interfacial tension, solubility and by the electrical charge on the molecules. Colloidal gold is inexpensive and preparation of a useful label is rapid, reproducible and the results easily quantitated from electron micrographs.

Staining of Concanavalin A-reactive glycoproteins on polyacrylamide gels with horseradish peroxidase — a critical evaluation

1984 ◽  
Vol 5 (2) ◽  
pp. 77-83 ◽  
Author(s):  
Klaus-Joachim Schott ◽  
Volker Neuhoff ◽  
Birgit Nessel ◽  
Ulla Pötter ◽  
Joachim Schröter
Keyword(s):  
Horseradish Peroxidase ◽  
Concanavalin A ◽  
Critical Evaluation ◽  
Polyacrylamide Gels

scholarly journals Peroxidase and gold complexes of lectins for double labeling of surface-binding sites by electron microscopy.

1977 ◽  
Vol 25 (10) ◽  
pp. 1181-1184 ◽  
Author(s):  
J Roth ◽  
M Wagner
Keyword(s):  
Electron Microscopy ◽  
Horseradish Peroxidase ◽  
Concanavalin A ◽  
Binding Sites ◽  
Lectin Binding ◽  
Helix Pomatia ◽  
Surface Binding ◽  
Double Labeling ◽  
Membrane Bound ◽  
Lectin Binding Sites

Double labeling experiments were performed for visualization of the binding sites of Concanavalin A and anti-AHel (the lectin from Helix pomatia). The anti-AHel was labeled with the colloidal gold whereas the membrane bound Concanavalin A was demonstrated by an affinity technique using horseradish peroxidase. The two markers used could be clearly distinguished electron microscopically. The specificity of the cell surface double labeling was demonstrated in the control experiments. A topological distinct localization of the both lectin-binding sites is evident.

scholarly journals The pinocytic rate of activated macrophages.

1975 ◽  
Vol 142 (5) ◽  
pp. 1150-1164 ◽  
Author(s):  
P J Edelson ◽  
R Zwiebel ◽  
Z A Cohn
Keyword(s):  
Horseradish Peroxidase ◽  
Concanavalin A ◽  
Peritoneal Macrophages ◽  
Intermediate Level ◽  
Cell Protein ◽  
Activated Macrophages ◽  
Peritoneal Cells ◽  
Activated State ◽  
Sustained Increase

Peritoneal macrophages from mice injected 4 days previously with Brewer's thioglycollate medium have a pinocytic rate, in culture, of 190 ng horseradish peroxidase (HRP)/100 mug cell protein/h, compared to the rate of resident peritoneal cells of 53 ng HRP/100 mug cell protein/h. Mice injected with endotoxin or with only certain of the components of the Brewer's medium show an intermediate level of stimulation. The rate of unstimulated, endotoxin-stimulated, or thioglycollate-stimulated cells shows little change over several days in culture. The pinocytic rate of thioglycollate-stimulated cells can, however, be further increased by exposure of concanavalin A. Although cells may show transient increases in their pinocytic rate in many situations, a sustained increase in pinocytic rate is a sign of the "activated" state of macrophages.

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