LCA recognizes sequences containing α-linked mannose residues but recognizes additional sugars as part of the receptor structure, giving it a narrower specificity than Con A. An α-linked fucose residue attached to the N-acetylchitobiose portion of the core oligosaccharide significantly enhances affinity. By exploiting this narrower specificity, glycoproteins and glycopeptides can be subfractionated with LCA after initial isolation with Con A.
Biotinylated Lens culinaris agglutinin 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.
| Unit Size | 5 mg |
|---|---|
| Applications | Immunohistochemistry / Immunocytochemistry, Immunofluorescence, Blotting Applications, Elispot, ELISAs, Glycobiology |
| Recommended Usage | For most applications we recommend a freshly prepared working solution of 5-20 µg/ml in the below buffer. |
| Recommended Storage | 2-8 °C; Store frozen for long term storage |
| Solution | 10 mM HEPES, 0.15 M NaCl, pH 7.5, 0.1 mM CaCl2, 0.01 mM MnCl2, 0.08% sodium azide, 10 mM methyl-alpha-D-mannopyranoside. |
| Concentration | 5 mg active conjugate/ml |
| Conjugate | Biotinylated |
| Sugar Specificity | Mannose, Glucose |
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 met. This may be particularly pertinent if using phosphate based buffers as diluents and storage solutions.
Lens culinaris agglutinin is composed of four subunits – two of about 17 kDa and two of 8 kDa. LCA has been found to be one of the most effective agents in preventing skin allograft rejection in model systems. LCA has been used to purify numerous glycoproteins, including immunoglobulins, histocompatibility antigens, and α2-macroglobulin.
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: mixture of 200 mM α-methylmannoside/200 mM α-methylglucoside
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