Lipid Metabolism
Adipogenesis and adipolysis are fundamental processes involved in fat metabolism. Understanding the chemical and physiological mechanisms behind the regulation of these two processes is an extremely active area of nutrition and health science research. Millipore offers two assays designed to aid researchers in the study of these vitally important physiological processes.
The Adipogenesis Assay provides a convenient format for induction and analysis of adipogenesis in the classic 3T3-L1 model. The most commonly used inducers of adipogenesis, dexamethasone, IBMX and insulin, are provided in convenient, ready-to-use formulations. Additionally, Oil Red O Solution, Wash Solution, and Dye Extraction Solution are included to stain and quantify lipid droplets in the differentiated adipocytes.
Adipogenesis Assay | |
3T3-L1 cells were plated in 24-well plates at 60,000 cells/well. The cells were incubated in DMEM containing calf serum alone (no inducers, left) or with IBMX and dexamethasone (+inducers, right) for 2 days. Cells were then incubated with the same media, except with insulin substituted for IBMX and dexamethasone for 2 days. Cells were then incubated with DMEM FCS for 3 days. Cells were stained with Oil Red O. |  |
Adipolysis Assay Kit
- Flexible format with components provided for use on a variety of culture plate sizes
- Quantitative and qualitative options for data or image analysis
The Adipolysis Assay Kit provides the necessary reagents for differentiating adipocytes and analyzing triglyceride mobilization by glycerol release. The kit includes IBMX Solution, Dexamethasone Solution, and Insulin Solution for addition to basal media (supplied by the user) to stimulate conversion of preadipocytes to lipid droplet-containing adipocytes. This assay kit is a convenient and sensitive tool for analysis of small molecules and proteins to stimulate or inhibit triglyceride breakdown in cultured adipocytes.
Kit Components
- Induction and Analysis
- Solutions
- Positive Controls
- Staining Solutions
Induction of adipolysis by â-adrenergic recepter activates the G-protein, Gás, that stimulates adenylate cyclase (AC) to produce cyclic AMP (cAMP). Protein Kinase A (PKA) is activated by cAMP to phosphorylate the lipid droplet surface protein perilipin (PL). Hormone-sensitive lipase (HSL) docks onto the phosphorylated PL and breaks down triglyceride into glycerol and free fatty acid. Glycerol is released into the extracellular space through aquaporin adipose (AQPad). |
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