Galanthus nivalis lectin, unlike most mannose-specific lectins, is not a metalloprotein and does not require Ca++ or Mn++ for binding.
Binding seems to be preferentially directed toward structures containing (α-1,3) mannose residues. Also in contrast to most mannose-binding lectins, GNL will not bind α-linked glucose. Reports indicate that this lectin binds rat and mouse IgM but not IgG. The only protein from human serum reported to bind to this lectin is α2-macroglobulin. GNL binds to many viral glycoproteins.
Biotinylated Galanthus nivalis lectin 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.
For most applications, we recommend a freshly prepared working solution of 5-20 µg/ml in the above buffer.
A precipitate may form during storage.This does not have a significant adverse effect
on the product.If a precipitate forms upon long-term storage, warm to 37 ºC and
centrifuge before use.
Recommended Storage
2-8 °C; Store frozen for long term storage
Solution
10 mM HEPES, pH 7.5, 0.15 M NaCl, 0.08% sodium azide, 0.1 mM CaCl2.
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.
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: 100 mM - 200 mM α-methylmannoside