Narcissus pseudonarcissus lectin or agglutinin (NPL/NPA) is affinity purified composed of four subunits. It agglutinates rabbit erythrocytes but is non-reactive with human erythrocytes. NPA has an isoelectric point between pH 4.2 and pH 4.6 with a specific binding activity toward α-D-mannose, although studies have suggested that the purified lectin may have greater affinity for internal mannosyl residues than for nonreducing terminal units. A buffer washed column elutes the sugar α-methylmannoside.
Human studies have found NPA to be nearly non-mitogenic, but it has been reported through an in vitro study that mitogenicity of NPA is age-related, with lymphoproliferative response of peripheral blood mononuclear cells (PBMC) in umbilical cord blood has a sevenfold increase than adult blood. NPA has also been found to be mitogenic for PBMC from lambs and not adult sheep.
Alkaline phosphatase (AP) is conjugated to Narcissus pseudonarcissus Lectin (NPL/NPA) to show the binding of NPL/NPA 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.