This lectin appears to bind only O-glycosidically linked oligosaccharides, preferring the structure galactosyl (β-1,3) N-acetylgalactosamine. This structure (the T-antigen) is the oligosaccharide to which peanut agglutinin (PNA) binds. However, unlike PNA, Jacalin will bind a mono- or disialylated form of this structure. This lectin has been used to purify human IgA. The specificity of this lectin also affords the opportunity to localize or isolate glycoproteins with O-glycosidically linked oligosaccharide side chains.
Biotinylated Jacalin has an appropriate number of biotins bound to provide the optimum staining characteristics for this lectin. This conjugate is supplied essentially free of unconjugated biotins and is preserved with sodium azide.
Reconstitute by adding 1 ml of water.Do not vortex.The resulting solution will have
the following composition:10 mM HEPES, 0.15 M NaCl, pH 7.5, 0.08% sodium azide,
0.1 mM CaCl2, 10 mM galactose, 10 mM lactose.
For most applications we recommend a freshly prepared working solution of 5-20 µg/ml
in the above buffer.
I recently purchased a biotinylated lectin. The datasheet supplied with the lectin suggests including 0.1 mM Ca++as part of the recommended buffer to prepare a working solution. What should I specifically add, and why is this required?
From our experience we have found that some lectins require Ca++ to be present for optimal binding activity. We suggest using calcium chloride (CaCl2) to fortify working solutions and ensure a minimum level of Ca++ is meet. This may be particularly pertinent if using phosphate based buffers as diluents and storage solutions.
Jacalin is a lectin composed of four subunits of approximately 16 kDa each.
This biotinylated lectin is an ideal intermediate for examining glycoconjugates using the Biotin-Avidin/Streptavidin System. First the biotinylated lectin is added, followed by the VECTASTAIN ABC Reagent, Avidin D conjugate, or streptavidin derivative.
Inhibiting/Eluting Sugar: 800 mM galactose or 100 mM melibiose