Assay for cell-bound lipase?

C.M. Longshaw MICCML at leeds.ac.uk
Thu Sep 18 08:29:40 EST 1997


Hi,
	I've been trying to find an method to assay lipase which 
may be either cell bound or cell associated and have had no real 
luck so far. The papers i have found suggest centrifugation, 
resuspension in 20mM HEPES and sonication, however i couldn't get 
any detectable activity from this method. Has anyone any other 
suggestions?

	The assay method i use at the momment is incubation with 
lipid emmulsion, extraction of fatty acids with dole reagent and 
titration of these using an auto-titrator. However i'm also 
interested in whether a spectrophotometric assay using 
p-nitrophenylpalmitate (pNPP) as a substrate might be easier (my 
supervisor has doubts whether this might be susceptible to esterase 
hydrolysis!). Any comments would be greatly appreciated,

Chris Longshaw,
Dep't Microbiology,
Leeds University
miccml at leeds.ac.uk
    



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