Affinity purification of natural ligands.
Immobilization of proteins, nucleic acids, and other "bioligands" requires selection of the proper bioligand for the specific application. This decision influences all others: the matrix is chosen and activated by the method that is most appropriate for this specific application and the ligand is coupled under conditions dictated by the activation method and the nature of the ligand (e.g., is it a labile protein or a sturdy enzyme cofactor?). There are many matrices and activation and coupling methods, and new ones are constantly being developed. This unit provides three protocols for activating and coupling proteins and other nucleophilic ligands to beaded agarose gel: cyanogen bromide, p-nitrophenyl chloroformate, and tresyl chloride.
Scouten
W H
WH
Utah State University, Logan, Utah, USA.
D002846:Chromatography, Affinity / Q000295:instrumentation / Q000379:methods*; D003488:Cyanogen Bromide / Q000737:chemistry; D008024:Ligands; D009696:Nucleic Acids / Q000737:chemistry; D011485:Protein Binding; D011506:Proteins / Q000737:chemistry*; D012685:Sepharose / Q000737:chemistry; D013450:Sulfones / Q000737:chemistry
D008024:Ligands; D009696:Nucleic Acids; D011506:Proteins; D013450:Sulfones; C032605:2,2,2-trifluoroethanesulfonyl chloride; D012685:Sepharose; D003488:Cyanogen Bromide