Lectins solution preparation

To remove excess lectin (particularly important if the 5-fold excess ratio is used), centrifuge the preparation at a minimum of 50,000g for 30 minutes to several hours (4°C), depending on the size of the particles and the amount of solution. Discard the supernatant, and resuspend the lectin-gold pellet in 0.01 M sodium phosphate, pH 7.4, containing 1 percent PEG. 

The interaction of partially purified, commercial preparations of P. vulgaris with a large number of serum glycoproteins was studied by Morse.105 The lectin precipitated a2-macroglobulin, /3-lipoproteins, and immunoglobulin M. Preparations ofarglycoprotein, orosomucoid, and several immunoglobulin A myeloma proteins also reacted.105 Chon-droitin 4-sulfate, dermatan sulfate, and heparin also precipitated a partially purified preparation of red kidney-bean the precipitation reaction was completely inhibited by 0.5 M sodium chloride solution.3663... 

For every protein purification problem there is always an affinity solution, but cost and safety considerations may render these solutions impractical. As an example, antibodies are widely used for analysis, where only relatively small amounts are usually required, but their production and purification on a large scale for preparative-scale chromatography may be difficult to justify economically. In some cases, Hhybridoma technology may be able to address this problem. Even if production costs are acceptable, the immobilized antibodies may be unstable over the sequence of sample application, elution, and sanitation required for multiple use of the affinity adsorbent. For these reasons, while biological ligands (antibodies, enzymes, receptors, lectin. 

Prepare glycan-binding protein solutions at desired concentrations in TSMBB. The GBPs are pre-labeled with either biotin or fluorescent dyes. We generally use biotinylated lectins at 10 pg/mL or lower (see Note 7). 

Glycoside-decorated amino-ether dendrons of up to third generation were prepared in solution and subsequently covalently immobilized on alumina chip by Pieters, Liskamp, and coworkers.The study of the binding of fluorescence-tagged lectins to the glycoclusters on the chip revealed the multivalency enhancement of the binding. 

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