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.
Horseradish peroxidase (HRP) is a 40 kDa protein that catalyzes the oxidation of substrates by hydrogen peroxide, resulting in a colored or fluorescent product or release of light as a byproduct of the reaction. It is most commonly used for blotting, immunoassays and immunohistochemistry methods. Narcissus pseudonarcissus Lectin (NPL/NPA) is conjugated to HRP at an appropriate ratio to provide optimal staining characteristics.