Cyanogen-bromide activated Sepharose

An immunoadsorbent has been prepared by attaching BSA, containing p-diazophenyl l-thio-/3-D-galactopyranosides, to cyanogen bromide-activated Sepharose.175 This affinity material was used to isolate mouse immunoglobulins (myeloma proteins) that bound carbohydrate.178... 

Sprinkle 1 g of cyanogen bromide-activated Sepharose 4B (Pharmacia-LKB, Piscataway, NJ) over 20 ml of 10 mM HC1. The gel swells immediately. One gram of dry gel yields 3.5 ml of hydrated matrix. 

Sepharose 4B, cyanogen bromide-activated-Sepharose, or protein A, G, or L Sepharose. 

Efficiencies less than 70% indicate that inefficient IgG immobilization has occurred. The cause of inefficient coupling is often the presence of a buffer component (free amines in Tris-HCl buffers, for example) that competes with primary amines of the antibody for binding at the active sites of the cyanogen bromide activated Sepharose beads. 

In another study, 5 -amino-C6 modified nucleic acid was immobilized onto cyanogen bromide activated Sepharose beads (Fig. 4B) [15]. The 100 pm-diameter beads were then subsequently transferred at the PDMS/air interface (Fig. 3D). 

A refined, affinity purification for L-fucose-binding Lotus proteins was reported by Blumberg and coworkers.684,688 N-(6-Amino-1-oxo-hexyl)-/3-L-fucopyranosylamine was coupled to cyanogen bromide-activated Sepharose 4B. The three proteins, A, B, and C, were bound by... 

Folate binding protein has been prepared from goat s milk by adsorption to a methotrexate-Sepharose column [94]. The folate binding protein was then coupled to cyanogen bromide activated Sepharose and used in the purification of folates and for the removal of endogenous folates from reaction mixtures. Cortisol hemisuccinate coupled to an amino Sepharose has been used to purify glucocorticoid binding protein(s) from rat liver... 

Recently Fallante et al. (200) reported that GSH S-transferase had been Imsioblllzed on a cyanogen bromide activated Sepharose gel, by a procedure similar to that discussed above for Immobilization of UDP-glucuronyltransferase (39), and used successfully for synthesis of GSH conjugates. A variety of substrates (Table VIII) were conjugated with this system and the products were characterized by chromatographic methods and FAB mass spectrometry. 

The attachment of amines on cyanogen bromide-activated Sepharose produces... 

Affinity chromatography. Concanavalin A was linked to cyanogen bromide activated Sepharose 4B by standard procedures (26). 

The remaining active groups on the cyanogen bromide activated Sepharose 4B were blocked by treatment with 1 M ethanolamine, pH 7.0 for 2 hr at 4°C. Approximately 90% of the concanavalin A was linked covalently to Sepharose 4B. Immobilized concanavalin A was activated by incubation overnight at 4°C in 40 mM Tris-HCl-40 NaCl-1 Ti MgCl2-l MnCl2-l tiM CaCl2. Before use the concanavalin A-Sepharose 4B preparation was equilibrated with 40 Tris-HCl-40 NaCl-0.24% DOC. 

Antibodies are affinity purified by binding them to immobilized antigen. The most common solid support for immobilizing antigen is cyanogen bromide-activated Sepharose. Almost all antigens containing amino... 

CNBr Activated Sepharose 4B - Cyanogen bromide-activated-Sepharose 4B... 

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