Lipase-based quantitation of triacylglycerols in cellular lipid extracts: requirement for presence of detergent and prior separation by thin-layer chromatography

Lipids
Paul P Van VeldhovenGuido Verhoeven

Abstract

A protocol, based on the use of Pseudomonas lipase, is presented to measure quantitatively the amount of triacylglycerols in extracts from cultured cells of tissues. Since the lipase also acts on di- and monoacylglycerols, separation of the extracts by thin-layer chromatography is recommended. In order to allow the lipase-catalyzed hydrolysis to proceed efficiently, lipid extracts or eluates from silica scrapings were mixed with the detergent Thesit [dodecylpoly(ethylene glycol ether)], prior to drying. After dissolution of the dried residues in water, the amount of triacylglycerols was quantified using Pseudomonas sp. lipase, glycerol kinase, glycerol-phosphate oxidase, and peroxidase. The activity of the latter enzyme was followed either colorimetrically in the presence of 4-aminoantipyrine and 2,4,6-tribromo-3-hydroxybenzoic acid or fluorimetrically in the presence of homovanillic acid.

References

Feb 1, 1967·Analytical Chemistry·G G GuilbualtE Hackley
Feb 1, 1984·Journal of Clinical Chemistry and Clinical Biochemistry. Zeitschrift Für Klinische Chemie Und Klinische Biochemie·U NägeleW Gruber
Jan 1, 1995·Journal of Inherited Metabolic Disease·P P Van Veldhoven

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Citations

Jul 7, 2007·Biochemistry and Cell Biology = Biochimie Et Biologie Cellulaire·Ewa PrzybytkowskiMarc Prentki
Jan 16, 2015·Scientific Reports·Joachim G SchulzCarlos G Dotti
Mar 19, 2014·Methods : a Companion to Methods in Enzymology·Jason M TennessenCarl S Thummel
Jun 22, 2019·Toxicology Mechanisms and Methods·Nibedita Nayak, Monalisa Mishra
Nov 1, 2016·Journal of Mammary Gland Biology and Neoplasia·Michael C RudolphPaul S MacLean
Nov 24, 2016·International Journal of Obesity : Journal of the International Association for the Study of Obesity·M C RudolphP S MacLean
Apr 9, 2020·Frontiers in Cell and Developmental Biology·Paul P Van VeldhovenElke Van Ael

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