Affinity purified Lens culinaris lectin (LCA) is composed of four subunits that consists of an α-chain (MW: 5.7 kDa) and β-chain (MW: 17.5 kDa). It has a carbohydrate specificity toward α-D-mannose and α-D-glucose, eluting with the sugar MeαMan+MeαGlc. This lectin is useful in affinity chromatography columns for the separation of glycoconjugates. The two isomers LCA-A and LCA-B agglutinates human red blood cells, although not blood group specific. Activity of LCA is determined by haemagglutination with human blood and agglutinates a 2% suspension of human erythrocytes at a lectin concentration of 8µg/ml. LCA requires calcium and manganese ions for binding. See other LCA conjugates.
Alkaline phosphatase (AP) is conjugated to Lens culinaris (Lentil) Lectin (LCA/LCH) to show the binding of LCA/LCH in many applications including Western blotting and ELISA. Alkaline phosphatase is a large protein (140 kDa) that catalyzes the hydrolysis of phosphate groups from a substrate resulting in a colored or fluorescent product. The optimal enzymatic activity of this protein is between pH 8 and 10, and its reaction rate remains linear, improving sensitivity over time.