What is CNBr activated Sepharose?
CNBr-activated Sepharose® 4B is a pre-activated medium for immobilization of ligands containing primary amines. It provides a very convenient way to immobilize ligands by the cyanogen bromide method. The coupling reaction is spontaneous, rapid and easy to carry out.
What is cyanogen bromide activated agarose?
Cyanogen bromide-activated Agarose is lyophilized powder stabilized with lactose used in affinity chromatography, protein chromatography, protein interactions, antibody labeling, antibody modification and attaching antibodies to agarose beads.
What is Sepharose 4B?
Sepharose® 4B is an agarose bead and a polymer of Gal β1,4 [3,6]-anhydro-L-galactose. Sepharose aids in fractionating larger molecules and is devoid of charged saccharides. Presence of 3,6-anhydro-L-galactose resists biological degradation. Hydrogen bonds stabilize the sepharose gel structure.
What are Sepharose beads?
Sepharose is a tradename for a crosslinked, beaded-form of agarose, a polysaccharide polymer material extracted from seaweed. Its brand name is a portmanteau derived from Separation-Pharmacia-Agarose. A common application for the material is in chromatographic separations of biomolecules.
What does cyanogen bromide do?
It is used in gold extraction, to make other chemicals, and as a fumigant. Cyanogen bromide (CNBr). A compound used in molecular biology to digest some proteins and as a coupling reagent for phosphoroamidate or pyrophosphate internucleotide bonds in DNA duplexes.
What are Sepharose beads used for?
Sepharose Big Beads: Purification from Crude, Viscous Samples at Large Scale. Use Sepharose Big Beads for purification of proteins from crude, viscous samples. Use Sepharose Big Beads when handling large volumes of crude or viscous samples that must be bound rapidly and when resolution is less important.
What does CNBr cleave?
Cyanogen bromide (CNBr) cleaves at methionine (Met) residues; BNPS-skatole cleaves at tryptophan (Trp) residues; formic acid cleaves at aspartic acid-proline (Asp-Pro) peptide bonds; hydroxylamine cleaves at asparagine-glycine (Asn-Gly) peptide bonds, and 2-nitro-5-thiocyanobenzoic acid (NTCB) cleaves at cysteine (Cys) …
What does CNBr stand for?
|CNBR||Chemical, Nuclear, Biological, and Radiation (weapons)|
|CNBR||Cyclic Nucleotide-Binding Region (physiology)|
What is Protein A Sepharose?
Protein A Sepharose® beads are prepared by covalently coupling recombinant Protein A to 6% cross-linked Sepharose® beads. Protein A is a genetically engineered protein containing five IgG-binding regions of native Protein A. This product can be used for IgG purification and immunoprecipitation.
What is crosslinked agarose?
Agarose is a very inert polysaccharide that forms hydrophilic and high gel strength gels at low concentrations. Cross-linked agarose beads are used in gel filtration chromatography (or molecular exclusion chromatography) as well as for activating beads for biomolecule purification or immobilization.
Are BrCN-sepharose beads stable in coupling buffer?
The BrCN-sepharose beads are NOT stable in coupling buffer. Let dry material swell in 10 ml 1 mM HCl for 15 min. 1 g material will give about 3.5 ml gel. Wash on a sintered glass filter (porosity G3) with about 200 ml 1 mM HCl per gram dry gel.
How to prepare the CnBr-activated Sepharose 4 b (Pharmacia)?
Preparation of the CNBr-activated Sepharose 4 B (Pharmacia) Start with these steps only when all materials are ready. The BrCN-sepharose beads are NOT stable in coupling buffer. Let dry material swell in 10 ml 1 mM HCl for 15 min. 1 g material will give about 3.5 ml gel.
What is the best buffer for protein coupling?
A buffer at pH 8.3 is most frequently used for coupling proteins. The high salt content of the coupling buffer minimizes protein-protein adsorption caused by the polyelectrolyte nature of proteins. Coupling of α-chymotrypsinogen by the method described here typically yields about 90% coupled protein.
What is the reaction between Cyanogen bromide and sepharose?
Cyanogen bromide reacts with hydroxyl groups on Sepharose to give a reactive product for coupling ligands via primary amino groups or similar nucleophilic groups. 1 Short term refers to the pH interval for regeneration, cleaning-in-place, and sanitization procedures.