The Con-A-peroxidase method for tissue localization of glucosyl and mannosyl groups applied to mouse hepatocytes and chicken erythrocytes.
A variation of the Concanavalin A (Con-A)-peroxidase labelling method originally described by Kiernan [Localization of alpha-D-glucosyl and alpha-D-mannosyl groups of mucosubstances with Concanavalin A and horseradish peroxidase. Histochemistry 1975;44:39-45] was applied to unsectioned cell preparations, with an emphasis on the nuclear localization of glycoproteins. Mouse liver imprints and chicken blood smears fixed in acetic acid-ethanol solution were studied. Modifications of the method included using increased Con-A concentration, and a range of pH values for the Con-A solutions. The strongest Con-A labelling of both erythrocytes and hepatocytes was obtained after incubation with Con-A at pH 6.5 and with Con-A concentrations at least two-fold greater than those used for tissue sections. These conditions may alter the Con-A conformation, enabling the lectin molecule to enter the cell nucleus and bind to nuclear glycoproteins, thus allowing their localization and quantification.
Moraes
Alberto S
AS
Department of Cell Biology, Institute of Biology, State University of Campinas (Unicamp), P.O. Box 6019, 13083-863, Campinas, São Paulo, Brazil.
Mello
Maria Luiza S
ML
D000818:Animals; D002645:Chickens; D003208:Concanavalin A / Q000737:chemistry*; D004900:Erythroblasts / Q000166:cytology / Q000378:metabolism*; D006023:Glycoproteins / Q000378:metabolism*; D022781:Hepatocytes / Q000166:cytology / Q000378:metabolism*; D006735:Horseradish Peroxidase / Q000737:chemistry; D007150:Immunohistochemistry / Q000379:methods; D008297:Male; D051379:Mice
D006023:Glycoproteins; D003208:Concanavalin A; D006735:Horseradish Peroxidase