The effect of n-butanol on Triton X-114 phase partitioning |
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Authors: | Ali Behdan L. Dale Snow |
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Affiliation: | (1) Biochemistry Laboratory, Carson-Taylor Hall, Louisiana Tech University, Ruston, Louisiana 71272, USA |
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Abstract: | n-Butanol interferes with the fractionation of amphiphilic and hydrophilic molecules during the Triton X-114 phase separation procedure. The indicators oil red (hydrophobic) and p-nitrophenol (hydrophilic) were useful for predicting the effectiveness of the Triton X-114 partition method. For n-butanol extracts containing oil red, 5-nucleotidase, or alkaline phosphatase, the hydrophobic molecules and Triton X-114 were retained in the aqueous phase during incubations at 30°C. The n-butanol interference was concentration-dependent and was reduced by lowering the final n-butanol concentration of the sample to 1.5% (v/v) or less. The results demonstrate how buffer-diluted n-butanol extracts of 5-nucleotidase and alkaline phosphatase can be successfully employed for subsequent Triton X-114 fractionation of the enzymes.This revised version was published online in October 2005 with corrections to the Cover Date. |
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Keywords: | alkaline phosphatase n-butanol extraction milk fat globule membrane p-nitrophenol 5 /content/w23415t744v78v8x/xxlarge8242.gif" alt=" prime" align=" BASELINE" BORDER=" 0" >-nucleotidase oil red Triton X-114 phase separation |
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