Concanavalin A is a lectin protein (MW 104kDa), homotetramer 26 kDa; originally extracted from the jack-bean, Canavalia ensiformis. It binds specifically to certain structures found in various sugars α-mannosyl and α-glucosyl residues in glycoproteins. It was the first lectin to be available on a commercial basis and is widely used in biology and biochemistry to characterize glycoproteins and other sugar-containing entities. It is also used to purify macromolecules in lectin affinity chromatography. Concanavalin A interacts with diverse receptors containing mannose carbohydrates (serum and membrane glycoproteins).ConA agglutinate strongly erythrocytes without being blood group specific, and cancerous cells. Normal cells agglutinate after trypsinisation. ConA is a also a lymphocyte mitogen.ConA reacts with many bacteria, like E. coli Dictyostelium discoideum et B. substilis It is also widely believed to be involved in the interaction between alpha-mannosyl oligosaccharides on the surface of the HIV virus and the human T cell lymphocyte.
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Histochemistry 1:250-1:1000; WB 1:500-1:2,500; ELISA 1:500-1:2,500.
Dilute biotinylated lectin in PBS containing 0.1 mM calcium chloride ions.
For Histochemistry: the tissues are processed same as for Immunohistochemistry, after blocking step biotinylated lectin is applied followed by streptavidin conjugated to enzyme and chromogen.
For WB: incubate membrane with blocking protein followed by biotinylated lectin, streptavidin enzyme conjugate and chromogen.
For ELISA: the target proteins are absorbed on ELISA plate at a concentration of 5-10 µg/ml, followed by blocking with protein solution followed by Biotinylated lectin, Streptavidin enzyme conjugate, ELISA substrate.
Please refer to Histochemistry, WB and ELISA protocol for detail information.
Please refer to Histochemistry, WB and ELISA protocol for detail information.