Please use this identifier to cite or link to this item:
|Title:||Application of isothermal titration calorimetry and column chromatography for identification of biomolecular targets||Authors:||Zhou, X.
|Issue Date:||Feb-2011||Citation:||Zhou, X., Kini, R.M., Sivaraman, J. (2011-02). Application of isothermal titration calorimetry and column chromatography for identification of biomolecular targets. Nature Protocols 6 (2) : 158-165. ScholarBank@NUS Repository. https://doi.org/10.1038/nprot.2010.187||Abstract:||This protocol describes a method for identifying unknown target proteins from a mixture of biomolecules for a given drug or a lead compound. This method is based on a combination of chromatography and isothermal titration calorimetry (ITC) where ITC is used as a tracking tool. The first step involves the use of ITC to confirm the binding of ligand to a component in the biomolecular mixture. Subsequently, the biomolecular mixture is fractionated by chromatography, and the binding of the ligand with individual fractions (or subfractions) is verified by ITC. The iteration of chromatographic purification on the fractions combined with ITC results in identifying the target protein. This method is useful when the target protein or ligand is unknown and/or not amenable to labeling, chemical modification or immobilization. This protocol has been successfully used by our team and by others to identify both low-abundance and highly abundant target proteins present in biomolecular mixtures. With this protocol, it takes approximately 3-5 d to identify the target protein from a mixture. © 2011 Nature America, Inc. All rights reserved.||Source Title:||Nature Protocols||URI:||http://scholarbank.nus.edu.sg/handle/10635/100106||ISSN:||17542189||DOI:||10.1038/nprot.2010.187|
|Appears in Collections:||Staff Publications|
Show full item record
Files in This Item:
There are no files associated with this item.
Items in DSpace are protected by copyright, with all rights reserved, unless otherwise indicated.