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Bioluminescent Assays For Measuring Steatosis and Insulin Action

Part # PS366

Abstract

Michael P. Valley, Natasha Karassina, and Jolanta Vidugiriene
Promega Corporation, 2800 Woods Hollow Rd, Madison, WI 53711

Nonalcoholic fatty liver disease (NAFLD), and its more serious form nonalcoholic steatohepatitis (NASH), are conditions in which lipids accumulate in the liver. To study these diseases, it is not only important to find good cellular models of steatosis, but it is also important to develop better assays to measure changes in lipid accumulation. Staining is routinely used to monitor lipid levels, but this is often nonspecific, somewhat laborious, and not quantitative. There are quantitative assays available, but most require an organic extraction, which is also undesirable.

We have developed a core bioluminescent technology that couples specific metabolite dehydrogenases to the production of NAD(P)H and the generation of light. These assays rely on detergents for lysis and extraction of lipids, hence, by utilizing specific dehydrogenases, lipases and esterases, we can quantify triglyceride, cholesterol and cholesterol esters without the requirement for an organic solvent.

Printed in USA.