The effect of n-butanol on Triton X-114 phase partitioning.
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 degrees 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.
Behdan
A
A
Biochemistry Laboratory, Louisiana Tech University, Ruston 71272, USA.
Snow
L D
LD
D020001:1-Butanol; D015720:5'-Nucleotidase / Q000302:isolation & purification*; D000469:Alkaline Phosphatase / Q000302:isolation & purification*; D000818:Animals; D001391:Azo Compounds; D002021:Buffers; D002417:Cattle; D006017:Glycolipids / Q000302:isolation & purification; D006023:Glycoproteins / Q000302:isolation & purification; D007202:Indicators and Reagents; D066292:Lipid Droplets; D008562:Membrane Glycoproteins / Q000302:isolation & purification; D009596:Nitrophenols; D017830:Octoxynol; D011092:Polyethylene Glycols; D014867:Water
D001391:Azo Compounds; D002021:Buffers; D006017:Glycolipids; D006023:Glycoproteins; D007202:Indicators and Reagents; D008562:Membrane Glycoproteins; D009596:Nitrophenols; C110367:milk fat globule; D014867:Water; D011092:Polyethylene Glycols; D020001:1-Butanol; D017830:Octoxynol; C010615:Nonidet P-40; D000469:Alkaline Phosphatase; D015720:5'-Nucleotidase; C011049:oil red O; C024836:4-nitrophenol