Tomato lectin, although sharing some specificities with potato lectin, Datura lectin, and wheat germ agglutinin, has been reported to be dissimilar in many respects. LEL binds well to glycophorin and Tamm-Horsfall glycoprotein and has been used effectively to label vascular endothelium in rodents.
Biotinylated tomato 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.
Can the biotinylated Lycopersicon esculentum (tomato) lectin be used for in vivo perfusion studies to trace blood vessels in mice?
The Lycopersicon esculentum (tomato) lectin has been widely reported as an effective blood vessel marker for in vivo vascular perfusion studies in rodent species. Investigators have primarily utilized one of the fluorophore conjugated tomato lectin formats to trace blood vasculature in normal and diseased animals via tail vein or intracardiac injection. However, the biotinylated format has also been used. It allows for flexibility in subsequent visualization by way of either fluorescence or enzyme-based methods. Published references are best source for procedural details. Examples of references where biotinylated tomato lectins have been applied via in vivo perfusion are featured below: Robertson, R.T., et al. (2014) Histochem. Cell Biol. 143(2) Thurston, G., et al. (1998) Am. J Pathol. 153(4):1099-1112
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.
Tomato lectin is a very stable single subunit glycoprotein containing about 50 percent arabinose and galactose and may form multimeric aggregates in solution.
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.