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Please use this identifier to cite or link to this item: http://eprint.iitd.ac.in/handle/2074/1371

Title: Purification of recombinant green fluorescent protein by three-phase partitioning
Authors: Jain, Sulakshana
Singh, Rajni
Gupta, M N
Keywords: Three-phase partitioning
Green fluorescent protein
Proteins
Issue Date: 2004
Citation: Journal of Chromatography A, 1035(1), 83-86
Abstract: The technique of three-phase partitioning (TPP) was used to purify the green fluorescent protein (GFP) in a single step. TPP uses a combination of ammonium sulphate and tert-butanol to precipitate proteins from their crude extracts. In the first round of TPP with 20% ammonium sulphate saturation at the ratio of crude to tert-butanol 1:1 (v/v), most of the GFP remains in the lower aqueous phase. When subjected to a second round of TPP with 60% ammonium sulphate saturation at the ratio of crude to tert-butanol 1:2 (v/v) gives 78% recovery of GFP with a 20-fold purification. The sodium dodecyl sulphate–polyacrylamide gel electrophoretic (SDS–PAGE) analysis of purified preparation shows single band. The fluorescence excitation and emission spectra agreed with values reported in literature.
URI: http://eprint.iitd.ac.in/dspace/handle/2074/1371
Appears in Collections:Chemistry

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