Lipogenesis Lipolysis Assay
Lipogenesis Lipolysis Assay - Related Videos
12:11
Determination of Fatty Acid Oxidation and Lipogenesis in Mouse Primary Hepatocytes
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Cited by 39 •
2015
De novo lipogenesis and β-fatty acid oxidation constitute key metabolic pathways in hepatocyte, pathways that are perturbed in several metabolic disorders, including fatty liver disease. Here we demonstrate isolation of mouse primary hepatocytes and describe quantification of β-fatty acid oxidation and lipogenesis.
07:59
Measurement of Basal and Forskolin-stimulated Lipolysis in Inguinal Adipose Fat Pads
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Cited by 10 •
2017
This protocol describes the method of determining basal and forskolin-stimulated lipolysis in inguinal fat pads obtained from normal chow diet (NCD) or high fat diet (HFD) ± capsaicin fed wild type mice. As an index for lipolysis, glycerol release was measured from inguinal adipose fat pads.
06:21
Hepatic Glucose Production, Ureagenesis, and Lipolysis Quantified using the Perfused Mouse Liver Model
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Cited by 4 •
2023
Here, we present a robust method for in situ perfusion of the mouse liver to study the acute and direct regulation of liver metabolism without disturbing the hepatic architecture but in the absence of extra-hepatic factors.
09:41
Measuring the Rate of Lipolysis in Ex Vivo Murine Adipose Tissue and Primary Preadipocytes Differentiated In Vitro
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Cited by 10 •
2023
Triglyceride lipolysis in adipocytes is an important metabolic process resulting in the liberation of free fatty acids and glycerol. Here, we provide a detailed protocol to measure basal and stimulated lipolysis in adipocytes and ex vivo adipose tissue from mice.
08:12
The TUNEL Assay
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2023
One of the hallmarks of apoptosis is the nuclear DNA fragmentation by nucleases. These enzymes are activated by caspases, the family of proteins that execute the cell death program. TUNEL assay is a method that takes advantage of this feature to detect apoptotic cells. In this assay, an enzyme called terminal deoxynucleotidyl transferase catalyzes the addition of dUTP nucleotides to the free 3’ ends of fragmented DNA. By using dUTPs that are labeled with chemical tags that can produce...
FAQs
The assay separates the two activities by selecting readouts that reflect their different biochemical directions. Lipogenesis can be assessed through lipid accumulation or incorporation of a substrate into cellular lipids, whereas lipolysis can be monitored through released glycerol and nonesterified fatty acids or changes in cellular lipid content. This distinction helps identify whether a treatment favors storage or mobilization.
Triacylglycerol hydrolysis produces glycerol and nonesterified fatty acids, so measuring these released products provides evidence of lipid breakdown. Their detection complements measurements of cellular lipid content, which may decrease as stored lipids are mobilized. Using product release alongside cellular measurements can therefore clarify whether an observed change reflects active lipolysis rather than simply reduced lipid formation.
Cellular state and experimental conditions strongly influence the measured balance between lipid formation and breakdown. Adipocyte differentiation affects the capacity for lipid storage, while hormonal responses can shift metabolic activity toward storage or mobilization. Defined conditions are important because changes in these variables may alter lipid accumulation, substrate incorporation, released products, or total cellular lipid content.
A suitable readout depends on the activity being examined and the information required. Researchers may track lipid accumulation, measure incorporation of a substrate into cellular lipids, quantify released glycerol or nonesterified fatty acids, or assess changes in total cellular lipid content. These complementary measurements can describe formation, breakdown, or the resulting change in stored lipid levels.
A general workflow applies defined experimental conditions to cells or biochemical samples, then measures a selected indicator of lipid formation or breakdown. Possible endpoints include accumulated cellular lipids, incorporated substrate, released hydrolysis products, or altered cellular lipid content. Comparing these endpoints under different conditions allows investigators to evaluate shifts in metabolic activity without treating storage and mobilization as identical processes.
The assay is useful when researchers need to examine metabolic regulation, adipocyte differentiation, or responses to hormones and potential treatments. It can reveal whether experimental conditions change lipid storage, mobilization, or both, providing evidence relevant to energy balance. In studies of obesity and related metabolic disorders, these measurements help evaluate how candidate interventions affect adipocyte lipid handling.