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Originally published In Press as doi:10.1194/jlr.D700021-JLR200 on September 13, 2007

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Journal of Lipid Research, Vol. 48, 2788-2791, December 2007
Copyright © 2007 by American Society for Biochemistry and Molecular Biology

Methods

Appraisal of hepatic lipase and lipoprotein lipase activities in mice

G. M. Dallinga-Thie1,*, A. J. Zonneveld-de Boer{dagger}, L. C. van Vark-van der Zee§, R. van Haperen**, T. van Gent**, H. Jansen{dagger}, R. De Crom** and A. van Tol**

* Department of Vascular Medicine, Academic Medical Center, Amsterdam, The Netherlands
{dagger} Department of Clinical Chemistry, Erasmus University Medical Center, Rotterdam, The Netherlands
§ Department of Vascular Medicine, Erasmus University Medical Center, Rotterdam, The Netherlands
** Department of Cell Biology and Genetics, Erasmus University Medical Center, Rotterdam, The Netherlands

Published, JLR Papers in Press, September 13, 2007.

1 To whom correspondence should be addressed. e-mail: g.m.dallinga{at}amc.uva.nl

A variety of methods are currently used to analyze HL and LPL activities in mice. In search of a simple methodology, we analyzed mouse preheparin and postheparin plasma LPL and HL activities using specific polyclonal antibodies raised in rabbit against rat HL (anti-HL) and in goat against rat LPL (anti-LPL). As an alternative, we analyzed HL activity in the presence of 1 M NaCl, a condition known to inhibit LPL activity in humans. The assays were validated using plasma samples from wild-type and HL-deficient C57BL/6 mice. We now show that the use of 1 M NaCl for the inhibition of plasma LPL activity in mice may generate incorrect measurements of both LPL and HL activities. Our data indicate that HL can be measured directly, without heparin injection, in preheparin plasma, because virtually all HL is present in an unbound form circulating in plasma. In contrast, measurable LPL activity is present only in postheparin plasma. Both HL and LPL can be measured using the same assay conditions (low salt and the presence of apolipoprotein C-II as an LPL activator). Total lipase activity in postheparin plasma minus preheparin HL activity reflects LPL activity. Specific antibodies are not required.

Supplementary key words high salt • preheparin plasma • postheparin plasma


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